TW202214286A - Anti-t cell antigen-binding molecule for use in combination with angiogenesis inhibitor - Google Patents

Abstract

The present disclosure provides a method for preventing, alleviating or treating the release of a cytokine or a side effect induced by the release of a cytokine by administering a VEGF inhibitor. For the purpose of preventing, alleviating or treating the release of a cytokine or a side effect induced by the release of a cytokine, a combination therapy using a lymphocyte-stimulating drug typified by an anti-T cell antigen-binding molecule and a VEGF inhibitor is also provided. For example, among anti-T cell antigen-binding molecules, an antibody capable of recruiting a T cell as an effector cell to a tumor tissue is also called "a T cell redirecting antibody" and is known as a means for treating a tumor. Meanwhile, when an antibody is bound to a T cell, if systemic cytokine production is stimulated, this systemic function may lead to disorders such as CRS. The present disclosure provides a means for reducing the systemic cytokine production, and therefore makes it possible to achieve the safer use of an anti-T cell antigen-binding molecule in the treatment of a tumor.

Description

Anti-T cell antigen binding molecules for use in combination with angiogenesis inhibitors

The present disclosure pertains to anti-T cell antigen binding molecules for use in combination with angiogenesis inhibitors. The present disclosure further relates to pharmaceutical compositions and methods for preventing, reducing, or treating side effects caused by cytokine production accompanying administration of anti-T cell antigen-binding molecules.

Antibodies are attracting attention because of their high stability in plasma and few side effects (Non-Patent Document 1 and Non-Patent Document 2). Antibody is known to induce not only the action of binding to the antigen, the action of agonist (agonist), the action of antagonism (antagonist), but also the induction of ADCC (Antibody Dependent Cytotoxicity: Antibody-dependent nociceptive activity), ADCP (Antibody Dependent Cell phagocytosis: Antibody Dependent Activity) The cytotoxic activity (also referred to as effector function) of effector cells, such as phagocytosis) and CDC (complement-dependent cytotoxic activity), exerts an antitumor effect on cancer cells (Non-Patent Document 3). So far, a plurality of therapeutic antibodies showing excellent antitumor effects have been developed as pharmaceuticals for the purpose of cancer treatment (Non-Patent Document 4). Although existing therapeutic antibodies are considered to have excellent effects, administration of such antibodies The therapeutic results obtained from antibodies still cannot meet the needs.

As molecules that inhibit multiple targets, antibodies (Bispecific antibodies, bispecific antibodies) that bind to two or more types of antigens in one molecule have been studied. If any of the antigens recognized by the bispecific antibody is an antigen that is specifically expressed in cancer, since it binds to any of the antigens, it shows cytotoxic activity against cancer cells, and it can be expected to be more than a normal antibody drug that recognizes a single antigen. More efficient anti-tumor effect.

As a kind of bispecific antibody, T cell redirecting antibody has been known since the 1980s, which is an antibody (T cell-redirecting antibody) that mobilizes T cells as effector cells and uses cell damage as a mechanism for its antitumor effect ( Non-patent documents 5, 6, 7). Unlike antibodies that mobilize NK cells, macrophages as effector cells, and ADCC as the mechanism for their antitumor effect, T cell redirecting antibodies are composed of T cell receptor (TCR) complexes directed against T cells An antibody of any of the units, in particular an antibody that binds to the CD3 epsilon chain, and a bispecific antibody that binds to an antigen on a target cancer cell. T cells approach cancer cells by binding T cell redirecting antibodies to both the CD3ε chain and the cancer antigen. As a result, it is considered that the antitumor effect on cancer cells is exerted by the cytotoxic effect of T cells. For example, in recent years, new T cell redirecting antibodies have been provided which have an Fc region in which two Fabs bind to cancer antigen (GPC3) and the CD3ε chain expressed on T cells, respectively, and have reduced FcγR-binding activity (Patent Document 1, 2).

On the other hand, it has been reported that the high antitumor activity exerted by T cell redirecting antibodies is accompanied by the occurrence of toxicity, for example, the occurrence of Cytokine Release Syndrome (CRS) (Non-Patent Document 8). [Prior Art Literature] [Patent Literature]

[Patent Document 1] WO2012/073985 (a concept patent of TRAB) [Patent Document 2] WO2016/047722 (ERY974 substance patent) [Non-patent literature]

[Non-Patent Document 1] Nat. Biotechnol. (2005) 23, 1073-1078 [Non-Patent Document 2] Eur J Pharm Biopharm. (2005) 59 (3), 389-396 [Non-Patent Document 3] Drug Des Devel Ther (2009) 3, 7-16 [Non-Patent Document 4] Clin Cancer Res. (2010) 16 (1), 11-20 [Non-Patent Document 5] Nature (1985) 314 (6012), 628-31 [Non-Patent Document 6] Int J Cancer (1988) 41 (4), 609-15. [Non-Patent Document 7] Proc Natl Acad Sci USA (1986) 83 (5), 1453-7 [Non-Patent Document 8] Cancer J. 2014 Mar-Apr; 20(2): 119-122.

[Problems to be Solved by Invention]

In a non-limiting aspect, the present disclosure relates to a combination therapy comprising an anti-T cell antigen-binding molecule and an angiogenesis inhibitor, a pharmaceutical composition used for the combination therapy, and the like. [Methods for solving problems]

In a non-limiting aspect, the inventors have discovered that administration of vascular endothelial growth factor (Vascular Endothelial Growth Factor; VEGF), a representative angiogenesis inhibitor, can prevent, alleviate, or treat concomitant disease in an individual. Cytokine-releasing syndrome (CRS) associated with administration of anti-T cell antigen-binding molecules.

In a non-limiting aspect, the present disclosure pertains to the following: (A1) A pharmaceutical composition comprising an anti-T cell antigen-binding molecule, wherein the pharmaceutical composition is used in combination with a vascular endothelial growth factor (VEGF) inhibitor to prevent and/or reduce and /or treatment of cytokine release syndrome and/or cytokine release. (A1-1) The pharmaceutical composition according to (A1), wherein the VEGF inhibitor is administered before, simultaneously with, or after the administration of the pharmaceutical composition. (A1-2) The pharmaceutical composition according to (A1), wherein the VEGF inhibitor is administered before or simultaneously with the administration of the pharmaceutical composition. (A1-3) The pharmaceutical composition according to any one of (A1) to (A1-2), which is administered 6 days, 5 days, 4 days, 3 days, The aforementioned VEGF inhibitor was administered 2 days before, 1 day before, or on the day before the administration of the aforementioned pharmaceutical composition. (A1-4) The pharmaceutical composition according to any one of (A1) to (A1-3), wherein the VEGF inhibitor is selected from an anti-VEGF antigen-binding molecule, an anti-VEGFR1 antigen-binding molecule, and an anti-VEGFR2 antigen-binding molecule , a VEGF receptor or a fusion protein comprising a fragment thereof, and more than one VEGF inhibitor of a tyrosine kinase inhibitor. (A1-5) The pharmaceutical composition according to any one of (A1) to (A1-4), wherein the VEGF inhibitor is selected from Bevacizumab, Ramucirumab, and one or more VEGF inhibitors of Aflibercept. (A1-6) The pharmaceutical composition according to any one of (A1) to (A1-5), which is a pharmaceutical composition for use in combination with an immune checkpoint inhibitor. (A1-7) The pharmaceutical composition according to (A1-6), wherein the immune checkpoint inhibitor is selected from the group consisting of anti-PD1 antibody, anti-PD-L1 antibody, and anti-PD-L2 antibody immune checkpoint inhibitor. (A1-8) The pharmaceutical composition according to (A1-6) or (A1-7), wherein the immune checkpoint inhibitor is atezolizumab. (A1-9) The pharmaceutical composition according to any one of (A1) to (A1-8), wherein no corticosteroid is administered before or simultaneously with the administration of the pharmaceutical composition. (A1-10) The pharmaceutical composition according to any one of (A1) to (A1-8), wherein a corticosteroid is further administered before, simultaneously with or after administration of the pharmaceutical composition. (A1-11) The pharmaceutical composition according to (A1-9) or (A1-10), wherein the corticosteroid is Dexamethasone, a pharmaceutically acceptable salt thereof, or a derivative thereof. (A1-12) The pharmaceutical composition according to any one of (A1) to (A1-11), wherein the anti-T cell antigen-binding molecule is a bispecific antigen-binding molecule comprising: (1) a domain comprising an antibody variable region having T cell receptor complex binding activity, and (2) A domain comprising an antibody variable region having cancer antigen-binding activity. (A1-13) The pharmaceutical composition according to any one of (A1) to (A1-12), wherein the anti-T cell antigen-binding molecule is a bispecific antibody comprising: (1) a domain comprising an antibody variable region having T cell receptor complex binding activity, (2) a domain comprising an antibody variable region having glypican3-binding activity, and (3) A domain comprising an Fc region whose binding activity to an Fcγ receptor is reduced. (A1-14) The pharmaceutical composition according to any one of (A1) to (A1-13), which is used for the treatment of cancer. (A1-15) The pharmaceutical composition according to any one of (A1) to (A1-14), which is used for preventing or reducing cytokine release syndrome and/or cytokine release, and treating cancer. (A1-16) The pharmaceutical composition according to any one of (A1) to (A1-15), wherein the cytokine release syndrome and/or cytokine release is accompanied by administration of the anti-T cell antigen-binding molecule Cytokine-releasing syndrome and/or cytokine release. (A1-17) The pharmaceutical composition according to any one of (A1) to (A1-16), wherein the cytokine release syndrome is a cytokine derived from either or both of non-tumor tissue and tumor tissue freed.

Or in a non-limiting aspect, the present disclosure is related to the following: (A2) A kit comprising (i) a container, (ii) a pharmaceutical composition containing an anti-T cell antigen-binding molecule within the container, and (iii) instructions prior to administration of the pharmaceutical composition, concurrently with Or after administration, for the prevention and/or alleviation and/or treatment of cytokine release syndrome and/or cytokine release, administration of a VEGF inhibitor. (A2-1) A kit comprising (i) a container, (ii) a pharmaceutical composition containing an anti-T cell antigen-binding molecule in the container, and (iii) indicating the administration of the aforementioned pharmaceutical composition 6 Days before, 5 days ago, 4 days ago, 3 days ago, 2 days ago, 1 day ago or on the same day, before the administration of the aforementioned pharmaceutical composition, in order to prevent and/or alleviate and/or treat cytokine release syndrome and Documentation of the release of cytokines for administration of VEGF inhibitors. (A2-2) The kit according to (A2) or (A2-1), wherein the aforementioned VEGF inhibitor is selected from anti-VEGF antigen-binding molecule, anti-VEGFR1 antigen-binding molecule, anti-VEGFR2 antigen-binding molecule, VEGF receptor or A fusion protein comprising a fragment thereof, and one or more VEGF inhibitors of tyrosine kinase inhibitors. (A2-3) The kit according to any one of (A2) to (A2-2), wherein the aforementioned VEGF inhibitor is one selected from bevacizumab, ramucirumab, and aflibercept The above VEGF inhibitors. (A2-4) The kit according to any one of (A2) to (A2-3), further comprising a label attached to a container showing that the pharmaceutical composition can be used for cancer treatment. (A2-5) The kit according to any one of (A2) to (A2-4), further comprising a document indicating that the aforementioned pharmaceutical composition is further used in combination with an immune checkpoint inhibitor. (A2-6) The kit according to (A2-5), wherein the immune checkpoint inhibitor is an immune checkpoint inhibitor selected from an anti-PD1 antibody, an anti-PD-L1 antibody, and an anti-PD-L2 antibody. (A2-7) The pharmaceutical composition according to (A2-5) or (A2-6), wherein the immune checkpoint inhibitor is atezolizumab. (A2-8) The kit according to any one of (A2) to (A2-7), further comprising a document indicating that corticosteroids are not administered before or at the same time as the pharmaceutical composition is administered . (A2-9) The kit according to any one of (A2) to (A2-7), further comprising a document indicating that corticosteroids are administered before, simultaneously with, or after administration of the aforementioned pharmaceutical composition . (A2-10) The kit according to (A2-8) or (A2-9), wherein the corticosteroid is Dexamethasone, a pharmaceutically acceptable salt thereof, or a derivative thereof. (A2-11) The kit according to any one of (A2) to (A2-10), wherein the anti-T cell antigen-binding molecule is a bispecific antigen-binding molecule comprising: (1) a domain comprising an antibody variable region having T cell receptor complex binding activity, and (2) A domain comprising an antibody variable region having cancer antigen-binding activity. (A2-12) The kit according to any one of (A2) to (A2-11), wherein the aforementioned anti-T cell antigen-binding molecule is a bispecific antibody comprising: (1) a domain comprising an antibody variable region having T cell receptor complex binding activity, (2) a domain comprising an antibody variable region having glypican3-binding activity, and (3) A domain comprising an Fc region whose binding activity to an Fcγ receptor is reduced. (A2-13) The kit according to any one of (A2) to (A2-12), wherein the prescription of the aforementioned pharmaceutical composition is carried out in accordance with the instructions of the aforementioned document. (A2-14) The kit according to any one of (A2) to (A2-13), for the treatment of cancer by the prescription indicated in the aforementioned document. (A2-15) The kit according to any one of (A2) to (A2-14), wherein the cytokine release syndrome and/or cytokine release are caused by administration of the anti-T cell antigen-binding molecule. Cytokine-releasing syndrome and/or cytokine release. (A2-16) The kit according to any one of (A2) to (A1-15), wherein the cytokine release syndrome is caused by the release of cytokines from either or both of the non-tumor tissue and the tumor tissue . (A2-17) The kit according to any one of (A2) to (A2-16), wherein the prescription of the aforementioned pharmaceutical composition is carried out in accordance with the instructions of the aforementioned document. (A3) Use of an anti-T cell antigen-binding molecule for the preparation of a pharmaceutical composition, characterized in that, by combined use with a VEGF inhibitor, it can prevent and/or alleviate and/or treat cytokine release syndrome and/or cytokines release, and treat cancer. (A3-1) The use according to (A3), wherein the VEGF inhibitor is administered before, simultaneously with, or after the administration of the pharmaceutical composition. (A3-2) The use according to (A3), wherein the VEGF inhibitor is administered before or simultaneously with the administration of the pharmaceutical composition. (A3-3) The use according to any one of (A3) to (A3-2), 6 days before, 5 days before, 4 days before, 3 days before, and 2 days before the administration of the pharmaceutical composition The aforementioned VEGF inhibitor is administered before, one day before, or on the same day, before the administration of the aforementioned pharmaceutical composition. (A3-4) The use according to any one of (A3) to (A3-3), wherein the VEGF inhibitor is selected from anti-VEGF antigen-binding molecule, anti-VEGFR1 antigen-binding molecule, anti-VEGFR2 antigen-binding molecule, VEGF A receptor or a fusion protein comprising a fragment thereof, and one or more VEGF inhibitors of tyrosine kinase inhibitors. (A3-5) The use according to any one of (A3) to (A3-4), wherein the VEGF inhibitor is one or more selected from bevacizumab, ramucirumab, and aflibercept VEGF inhibitor. (A3-6) The use according to any one of (A3) to (A3-5), wherein the pharmaceutical composition is further used in combination with an immune checkpoint inhibitor. (A3-7) The use according to (A3-6), wherein the immune checkpoint inhibitor is an immune checkpoint inhibitor selected from an anti-PD1 antibody, an anti-PD-L1 antibody, and an anti-PD-L2 antibody. (A3-8) The use according to (A3-6) or (A3-7), wherein the immune checkpoint inhibitor is atezolizumab. (A3-9) The use according to any one of (A3) to (A3-8), wherein no corticosteroid is administered before or simultaneously with the administration of the pharmaceutical composition. (A3-10) The use according to any one of (A3) to (A3-8), wherein a corticosteroid is further administered before, simultaneously with, or after the administration of the pharmaceutical composition. (A3-11) The use according to (A3-9) or (A3-10), wherein the corticosteroid is Dexamethasone, a pharmaceutically acceptable salt thereof, or a derivative thereof. (A3-12) The use according to any one of (A3) to (A3-11), wherein the anti-T cell antigen-binding molecule is a bispecific antigen-binding molecule comprising: (1) a domain comprising an antibody variable region having T cell receptor complex binding activity, and (2) A domain comprising an antibody variable region having cancer antigen-binding activity. (A3-13) The use according to any one of (A3) to (A3-12), wherein the anti-T cell antigen-binding molecule is a bispecific antibody comprising: (1) a domain comprising an antibody variable region having T cell receptor complex binding activity, (2) a domain comprising an antibody variable region having glypican3-binding activity, and (3) A domain comprising an Fc region whose binding activity to an Fcγ receptor is reduced. (A3-14) The use according to any one of (A3) to (A3-13), wherein the cytokine release syndrome and/or cytokine release is caused by administration of the anti-T cell antigen-binding molecule Cytokine Release Syndrome and/or Cytokine Release. (A3-15) The use according to any one of (A3) to (A3-14), wherein the cytokine release syndrome is caused by the release of cytokines in either or both of non-tumor tissues and tumor tissues. (A4) Use of an anti-T cell antigen-binding molecule and a VEGF inhibitor in combination therapy for preventing and/or alleviating and/or treating cytokine release syndrome and/or cytokine release, and treating cancer. (A4-1) The use according to (A4), wherein the VEGF inhibitor is administered before, simultaneously with, or after the administration of the anti-T cell antigen-binding molecule in the combination therapy. (A4-2) The use according to (A4), wherein the VEGF inhibitor is administered before or simultaneously with the administration of the anti-T cell antigen-binding molecule in the combination therapy. (A4-3) The use according to any one of (A4) to (A4-2), in the combination therapy, 6 days before, 5 days before, 4 days before the administration of the aforementioned anti-T cell antigen-binding molecule The aforementioned VEGF inhibitor is administered 1 day before, 3 days ago, 2 days ago, 1 day ago, or on the day before the administration of the aforementioned anti-T cell antigen-binding molecule. (A4-4) The use according to any one of (A4) to (A4-3), wherein the VEGF inhibitor is selected from anti-VEGF antigen-binding molecule, anti-VEGFR1 antigen-binding molecule, anti-VEGFR2 antigen-binding molecule, VEGF A receptor or a fusion protein comprising a fragment thereof, and one or more VEGF inhibitors of tyrosine kinase inhibitors. (A4-5) The use according to any one of (A4) to (A4-4), wherein the VEGF inhibitor is one or more selected from bevacizumab, ramucirumab, and aflibercept VEGF inhibitor. (A4-6) The use according to any one of (A4) to (A4-5), wherein the combination therapy is further administered with an immune checkpoint inhibitor. (A4-7) The use according to (A4-6), wherein the immune checkpoint inhibitor is an immune checkpoint inhibitor selected from an anti-PD1 antibody, an anti-PD-L1 antibody, and an anti-PD-L2 antibody. (A4-8) The use according to (A4-6) or (A4-7), wherein the aforementioned immune checkpoint inhibitor is Atezolizumab. (A4-9) The use according to any one of (A4) to (A4-8), wherein in the combination therapy, no corticosteroid is administered before or simultaneously with the administration of the anti-T cell antigen-binding molecule (Corticosteroid). (A4-10) The use according to any one of (A4) to (A4-8), wherein in the combined therapy, the anti-T cell antigen-binding molecule is administered before, at the same time, or after the administration. Give corticosteroids. (A4-11) The use according to (A4-9) or (A4-10), wherein the corticosteroid is Dexamethasone, a pharmaceutically acceptable salt thereof, or a derivative thereof. (A4-12) The use according to any one of (A4) to (A4-11), wherein the anti-T cell antigen-binding molecule is a bispecific antigen-binding molecule comprising: (1) a domain comprising an antibody variable region having T cell receptor complex binding activity, and (2) A domain comprising an antibody variable region having cancer antigen-binding activity. (A4-13) The use according to any one of (A4) to (A4-12), wherein the anti-T cell antigen-binding molecule is a bispecific antibody comprising: (1) a domain comprising an antibody variable region having T cell receptor complex binding activity, (2) a domain comprising an antibody variable region having glypican3-binding activity, and (3) A domain comprising an Fc region whose binding activity to an Fcγ receptor is reduced. (A4-14) The use according to any one of (A4) to (A4-13), wherein the cytokine-releasing syndrome and/or cytokine release are cells accompanying the administration of the anti-T cell antigen-binding molecule Hormone release syndrome and/or cytokine release. (A4-15) The use according to any one of (A4) to (A4-14), wherein the cytokine release syndrome is caused by the release of cytokines in either or both of non-tumor tissues and tumor tissues.

Alternatively, in a non-limiting aspect, the present disclosure pertains to: (A5) a method of treating cancer, preventing and/or alleviating and/or treating cytokine release syndrome and/or cytokine release by administering an anti-T cell antigen-binding molecule to a subject in combination with a VEGF inhibitor, and treat cancer. (A5-1) The method according to (A5), wherein the VEGF inhibitor is administered before, simultaneously with, or after the administration of the anti-T cell antigen-binding molecule. (A5-2) The method according to (A5), wherein the VEGF inhibitor is administered before or simultaneously with the administration of the anti-T cell antigen-binding molecule. (A5-3) The method according to any one of (A5) to (A5-2), 6 days, 5 days, 4 days, and 3 days before the administration of the anti-T cell antigen-binding molecule , 2 days before, 1 day before, or on the same day, before the administration of the aforementioned anti-T cell antigen-binding molecule, the aforementioned VEGF inhibitor was administered. (A5-4) The method according to any one of (A5) to (A5-3), wherein the VEGF inhibitor is selected from anti-VEGF antigen-binding molecules, anti-VEGFR1 antigen-binding molecules, anti-VEGFR2 antigen-binding molecules, VEGF A receptor or a fusion protein comprising a fragment thereof, and one or more VEGF inhibitors of tyrosine kinase inhibitors. (A5-5) The method according to any one of (A5) to (A5-4), wherein the VEGF inhibitor is one or more selected from bevacizumab, ramucirumab, and aflibercept VEGF inhibitor. (A5-6) The method according to any one of (A5) to (A5-5), further comprising using it together with an immune checkpoint inhibitor. (A5-7) The method according to (A5-6), wherein the immune checkpoint inhibitor is an immune checkpoint inhibitor selected from an anti-PD1 antibody, an anti-PD-L1 antibody, and an anti-PD-L2 antibody. (A5-8) The method according to (A5-6) or (A5-7), wherein the immune checkpoint inhibitor is atezolizumab. (A5-9) The method according to any one of (A5) to (A5-8), wherein no corticosteroid is administered before or simultaneously with the administration of the anti-T cell antigen-binding molecule. (A5-10) The method according to any one of (A5) to (A5-8), wherein a corticosteroid is further administered before, simultaneously with, or after the administration of the anti-T cell antigen-binding molecule. (A5-11) The method according to (A5-9) or (A5-10), wherein the corticosteroid is dexamethasone, a pharmaceutically acceptable salt thereof, or a derivative thereof. (A5-12) The method according to any one of (A5) to (A5-11), wherein the anti-T cell antigen-binding molecule is a bispecific antigen-binding molecule comprising: (1) a domain comprising an antibody variable region having T cell receptor complex binding activity, and (2) A domain comprising an antibody variable region having cancer antigen-binding activity. (A5-13) The method according to any one of (A5) to (A5-12), wherein the anti-T cell antigen-binding molecule is a bispecific antibody comprising: (1) a domain comprising an antibody variable region having T cell receptor complex binding activity, (2) a domain comprising an antibody variable region having glypican3-binding activity, and (3) A domain comprising an Fc region whose binding activity to an Fcγ receptor is reduced. (A5-14) The method according to any one of (A5) to (A5-13), wherein the cytokine release syndrome and/or cytokine release is a cell accompanied by the administration of the anti-T cell antigen-binding molecule Hormone release syndrome and/or cytokine release. (A5-15) As in (A) the aforementioned cytokine release syndrome is caused by the release of cytokines in either or both of the non-tumor tissue and the tumor tissue.

Or this disclosure includes the following aspects: (B1) A pharmaceutical composition for preventing and/or alleviating and/or treating cytokine release syndrome and/or cytokine release, comprising a VEGF inhibitor. (B1-1) The pharmaceutical composition according to (B1), wherein the VEGF inhibitor is selected from an anti-VEGF antigen-binding molecule, an anti-VEGFR1 antigen-binding molecule, an anti-VEGFR2 antigen-binding molecule, a VEGF receptor, or a fusion comprising a fragment thereof A protein, and one or more VEGF inhibitors of tyrosine kinase inhibitors. (B1-2) The pharmaceutical composition according to (B1) or (B1-1), wherein the VEGF inhibitor is selected from the group consisting of bevacizumab, ramucirumab, and aflibercept. (B1-3) The pharmaceutical composition according to any one of (B1) to (B1-2), which is a pharmaceutical composition for combined use with an immune checkpoint inhibitor. (B1-4) The pharmaceutical composition according to (B1-3), wherein the immune checkpoint inhibitor is an immune checkpoint inhibitor selected from an anti-PD1 antibody, an anti-PD-L1 antibody, and an anti-PD-L2 antibody. (B1-5) The pharmaceutical composition according to (B1-3) or (B1-4), wherein the immune checkpoint inhibitor is atezolizumab. (B1-6) The pharmaceutical composition according to any one of (B1) to (B1-5), for use in combination therapy with an anti-T cell antigen-binding molecule. (B1-7) The pharmaceutical composition according to (B1-6), which is used for administration before, simultaneously with, or after administration of the anti-T cell antigen-binding molecule. (B1-8) The pharmaceutical composition according to (B1-6), which is used for administration before or simultaneously with the administration of the anti-T cell antigen-binding molecule. (B1-9) The pharmaceutical composition according to any one of (B1-6) to (B1-8), which is 6 days, 5 days, or 4 days before the administration of the anti-T cell antigen-binding molecule , 3 days ago, 2 days ago, 1 day ago, or the same day, for administration before the administration of the aforementioned anti-T cell antigen-binding molecule. (B1-10) The pharmaceutical composition according to any one of (B1-6) to (B1-9), wherein no corticosteroid is administered before or simultaneously with the administration of the anti-T cell antigen-binding molecule ). (B1-11) The pharmaceutical composition according to any one of (B1-6) to (B1-9), which is further administered to the cortex before, simultaneously with, or after the administration of the anti-T cell antigen-binding molecule. Steroid. (B1-12) The pharmaceutical composition according to (B1-10) or (B1-11), wherein the corticosteroid is Dexamethasone, a pharmaceutically acceptable salt thereof, or a derivative thereof. (B1-13) The pharmaceutical composition according to any one of (B1-6) to (B1-12), wherein the anti-T cell antigen-binding molecule is a bispecific antigen-binding molecule comprising: (1) a domain comprising an antibody variable region having T cell receptor complex binding activity, and (2) A domain comprising an antibody variable region having cancer antigen-binding activity. (B1-14) The pharmaceutical composition according to any one of (B1-6) to (B1-13), wherein the anti-T cell antigen-binding molecule is a bispecific antibody comprising: (1) a domain comprising an antibody variable region having T cell receptor complex binding activity, (2) a domain comprising an antibody variable region having glypican3-binding activity, and (3) A domain comprising an Fc region whose binding activity to an Fcγ receptor is reduced. (B1-15) The pharmaceutical composition according to any one of (B1-6) to (B1-14), wherein the combination of the pharmaceutical composition and the anti-T cell antigen-binding molecule is used for prophylaxis and/or Combination of alleviating and/or treating cytokine release syndrome and/or cytokine release and treating cancer. (B1-16) The pharmaceutical composition according to any one of (B1-6) to (B1-15), wherein the cytokine release syndrome and/or cytokine release is accompanied by the anti-T cell antigen-binding molecule Administered cytokine release syndrome and/or cytokine release. (B1-17) The pharmaceutical composition according to any one of (B1) to (B1-16), wherein the cytokine release syndrome is a cytokine derived from either or both of non-tumor tissue and tumor tissue freed.

Or in a non-limiting aspect, the present disclosure is related to the following: (B2) A kit comprising (i) a container, (ii) a pharmaceutical composition comprising a VEGF inhibitor in the container, and (iii) for preventing and/or alleviating and/or treating cytokine release syndrome and/or Cytokine release, documentation indicating administration of the pharmaceutical composition. (B2-1) The kit according to (B2), wherein the VEGF inhibitor is selected from anti-VEGF antigen-binding molecules, anti-VEGFR1 antigen-binding molecules, anti-VEGFR2 antigen-binding molecules, VEGF receptors, or fusion proteins comprising fragments thereof , and one or more cytokine inhibitors of tyrosine kinase inhibitors. (B2-2) The kit according to (B2) or (B2-1), wherein the VEGF inhibitor is one or more VEGF inhibitor selected from bevacizumab, ramucirumab, and aflibercept agent. (B2-3) The kit according to (B2) to (B2-2), further comprising a document indicating the combined use of the aforementioned pharmaceutical composition and an immune checkpoint inhibitor. (B2-4) The kit according to (B2-3), wherein the immune checkpoint inhibitor is an immune checkpoint inhibitor selected from an anti-PD1 antibody, an anti-PD-L1 antibody, and an anti-PD-L2 antibody. (B2-5) The pharmaceutical composition according to (B2-3) or (B2-4), wherein the immune checkpoint inhibitor is atezolizumab. (B2-6) The kit according to any one of (B2) to (B2-5), further comprising a document indicating that the aforementioned pharmaceutical composition is used in combination with an anti-T cell antigen-binding molecule. (B2-7) The kit according to (B2-6), further comprising a document instructing to administer the pharmaceutical composition before, simultaneously with, or after the administration of the anti-T cell antigen-binding molecule. (B2-8) The kit according to (B2-6), further comprising a document instructing to administer the pharmaceutical composition before or simultaneously with the administration of the anti-T cell antigen-binding molecule. (B2-9) The kit according to any one of (B2-6) to (B2-8), further comprising instructing the pharmaceutical composition to be administered 6 days before the administration of the anti-T cell antigen-binding molecule, 5 days ago, 4 days ago, 3 days ago, 2 days ago, 1 day ago, or the day before the administration of the aforementioned anti-T cell antigen-binding molecule. (B2-10) The kit according to any one of (B2-6) to (B2-9), further comprising an instruction to not administer corticosteroids before or simultaneously with the administration of the aforementioned anti-T cell antigen-binding molecule (Corticosteroid). (B2-11) The kit according to any one of (B2-6) to (B2-9), further comprising an instruction to further administer the anti-T cell antigen-binding molecule before, at the same time, or after the administration. File for corticosteroids. (B2-12) The kit according to (B2-10) or (B2-11), wherein the corticosteroid is Dexamethasone, a pharmaceutically acceptable salt thereof, or a derivative thereof. (B2-13) The kit according to any one of (B2-6) to (B2-12), wherein the anti-T cell antigen-binding molecule is a bispecific antigen-binding molecule comprising: (1) a domain comprising an antibody variable region having T cell receptor complex binding activity, and (2) A domain comprising an antibody variable region having cancer antigen-binding activity. (B2-14) The kit according to any one of (B2-6) to (B2-13), wherein the anti-T cell antigen-binding molecule is a bispecific antibody comprising: (1) a domain comprising an antibody variable region having T cell receptor complex binding activity, (2) a domain comprising an antibody variable region having glypican3-binding activity, and, (3) A domain comprising an Fc region whose binding activity to an Fcγ receptor is reduced. (B2-15) The kit according to any one of (B2-6) to (B2-14), wherein the combined use of the aforementioned pharmaceutical composition and the aforementioned anti-T cell antigen-binding molecule is for preventing and/or reducing And/or treatment of cytokine release syndrome and/or cytokine release, and the combination of treatment of cancer. (B2-16) The kit according to any one of (B2-6) to (B2-15), wherein the cytokine release syndrome and/or cytokine release is accompanied by administration of the anti-T cell antigen-binding molecule Cytokine release syndrome and/or cytokine release. (B2-17) The kit according to any one of (B2) to (B2-16), wherein the cytokine release syndrome is caused by the release of cytokines from either or both of the non-tumor tissue and the tumor tissue . (B2-18) The kit according to any one of (B2) to (B2-17), wherein the prescription of the aforementioned pharmaceutical composition is performed in accordance with the instructions of the aforementioned document. (B3) Use of a VEGF inhibitor in the preparation of a pharmaceutical composition for preventing and/or alleviating and/or treating cytokine release syndrome and/or cytokine release. (B3-1) The use according to (B3), wherein the aforementioned VEGF inhibitor is selected from anti-VEGF antigen-binding molecules, anti-VEGFR1 antigen-binding molecules, anti-VEGFR2 antigen-binding molecules, VEGF receptors or fusion proteins comprising fragments thereof, and one or more cytokine inhibitors of tyrosine kinase inhibitors. (B3-2) The use according to (B3) or (B3-1), wherein the aforementioned VEGF inhibitor is one or more VEGF inhibitors selected from bevacizumab, ramucirumab, and aflibercept . (B3-3) The use according to any one of (B3) to (B3-2), wherein the pharmaceutical composition is a pharmaceutical composition used in combination with an immune checkpoint inhibitor. (B3-4) The use according to (B3-3), wherein the immune checkpoint inhibitor is an immune checkpoint inhibitor selected from an anti-PD1 antibody, an anti-PD-L1 antibody, and an anti-PD-L2 antibody. (B3-5) The use according to (B3-3) or (B3-4), wherein the aforementioned immune checkpoint inhibitor is atezolizumab. (B3-6) The use according to any one of (B3) to (B3-5), wherein the pharmaceutical composition is a pharmaceutical composition for combination therapy with an anti-T cell antigen-binding molecule. (B3-7) The use according to (B3-6), wherein the pharmaceutical composition is administered before, simultaneously with, or after the administration of the anti-T cell antigen-binding molecule. (B3-8) The use according to (B3-6), wherein the pharmaceutical composition is administered before or simultaneously with the administration of the anti-T cell antigen-binding molecule. (B3-9) The use according to any one of (B3-6) to (B3-8), 6 days before, 5 days before, 4 days before, 3 days before the administration of the anti-T cell antigen-binding molecule The pharmaceutical composition is administered 1 day before, 2 days ago, 1 day before, or on the same day before the administration of the aforementioned anti-T cell antigen-binding molecule. (B3-10) The use according to any one of (B3-6) to (B3-9), wherein no corticosteroid is administered before or simultaneously with the administration of the anti-T cell antigen-binding molecule. (B3-11) The use according to any one of (B3-6) to (B3-9), wherein a corticosteroid is further administered before, simultaneously with or after the administration of the anti-T cell antigen-binding molecule. (B3-12) The use according to (B3-10) or (B3-11), wherein the corticosteroid is Dexamethasone, a pharmaceutically acceptable salt thereof, or a derivative thereof. (B3-13) The use according to any one of (B3-6) to (B3-12), wherein the anti-T cell antigen-binding molecule is a bispecific antigen-binding molecule comprising: (1) a domain comprising an antibody variable region having T cell receptor complex binding activity, and (2) A domain comprising an antibody variable region having cancer antigen-binding activity. (B3-14) The use according to any one of (B3-6) to (B3-13), wherein the anti-T cell antigen-binding molecule is a bispecific antibody comprising: (1) a domain comprising an antibody variable region having T cell receptor complex binding activity, (2) a domain comprising an antibody variable region having glypican3-binding activity, and, (3) A domain comprising an Fc region whose binding activity to an Fcγ receptor is reduced. (B3-15) The use according to any one of (B3-6) to (B3-14), wherein the combination therapy of the aforementioned pharmaceutical composition and the aforementioned anti-T cell antigen-binding molecule is for prevention and/or alleviation and Combination therapy for the treatment of cytokine release syndrome and/or cytokine release, and the treatment of cancer. (B3-16) The use according to any one of (B3-6) to (B3-15), wherein the cytokine release syndrome and/or cytokine release is accompanied by administration of the anti-T cell antigen-binding molecule Cytokine Release Syndrome and/or Cytokine Release. (B3-17) The use according to any one of (B3) to (B3-16), wherein the cytokine release syndrome is caused by the release of cytokines in either or both of the non-tumor tissue and the tumor tissue. (B4) Use of a VEGF inhibitor for preventing and/or alleviating and/or treating cytokine release syndrome and/or cytokine release. (B4-1) The use according to (B4), wherein the aforementioned VEGF inhibitor is selected from anti-VEGF antigen-binding molecules, anti-VEGFR1 antigen-binding molecules, anti-VEGFR2 antigen-binding molecules, VEGF receptors or fusion proteins comprising fragments thereof, and one or more VEGF inhibitors of tyrosine kinase inhibitors. (B4-2) The use according to (B4) or (B4-1), wherein the aforementioned VEGF inhibitor is one or more VEGF inhibitors selected from bevacizumab, ramucirumab, and aflibercept . (B4-3) The use according to any one of (B4) to (B4-2), wherein the aforementioned VEGF inhibitor is used in combination with an immune checkpoint inhibitor. (B4-4) The use according to (B4-3), wherein the immune checkpoint inhibitor is an immune checkpoint inhibitor selected from an anti-PD1 antibody, an anti-PD-L1 antibody, and an anti-PD-L2 antibody. (B4-5) The use according to (B4-3) or (B4-4), wherein the aforementioned immune checkpoint inhibitor is atezolizumab. (B4-6) The use according to any one of (B4) to (B4-5), wherein the aforementioned VEGF inhibitor is used in combination with an anti-T cell antigen-binding molecule. (B4-7) The use according to (B4-6), wherein the VEGF inhibitor is administered before, simultaneously with, or after the administration of the anti-T cell antigen-binding molecule. (B4-8) The use according to (B4-6), wherein the VEGF inhibitor is administered before or simultaneously with the administration of the anti-T cell antigen-binding molecule. (B4-9) The use according to any one of (B4-6) to (B4-8), wherein the VEGF inhibitor is administered 6 days before, 5 days before, or 5 days before the administration of the anti-T cell antigen-binding molecule. Administration was performed 4 days before, 3 days before, 2 days before, 1 day before, or on the day before administration of the aforementioned anti-T cell antigen-binding molecule. (B4-10) The use according to any one of (B4-6) to (B4-9), wherein no corticosteroid is administered before or simultaneously with the administration of the anti-T cell antigen-binding molecule. (B4-11) The use according to any one of (B4-6) to (B4-9), wherein a corticosteroid is further administered before, simultaneously with or after the administration of the anti-T cell antigen-binding molecule. (B4-12) The use according to (B4-10) or (B4-11), wherein the aforementioned corticosteroid is Dexamethasone, a pharmaceutically acceptable salt thereof, or a derivative thereof. (B4-13) The use according to any one of (B4-6) to (B4-12), wherein the anti-T cell antigen-binding molecule is a bispecific antigen-binding molecule comprising: (1) a domain comprising an antibody variable region having T cell receptor complex binding activity, and (2) A domain comprising an antibody variable region having cancer antigen-binding activity. (B4-14) The use according to any one of (B4-6) to (B4-13), wherein the anti-T cell antigen-binding molecule is a bispecific antibody comprising: (1) a domain comprising an antibody variable region having T cell receptor complex binding activity, (2) a domain comprising an antibody variable region having glypican3-binding activity, and, (3) A domain comprising an Fc region whose binding activity to an Fcγ receptor is reduced. (B4-15) The use according to any one of (B4-6) to (B4-14), wherein the combined use of the aforementioned VEGF inhibitor and the aforementioned anti-T cell antigen-binding molecule is for prevention and/or mitigation and/or Or the combination of treating cytokine release syndrome and/or cytokine release and treating cancer. (B4-16) The use according to any one of (B4-6) to (B4-15), wherein the cytokine release syndrome and/or cytokine release is accompanied by administration of the anti-T cell antigen-binding molecule Cytokine Release Syndrome and/or Cytokine Release. (B4-17) The use according to any one of (B4) to (B4-16), wherein the cytokine release syndrome is caused by the release of cytokines in either or both of the non-tumor tissue and the tumor tissue.

Alternatively, in a non-limiting aspect, the present disclosure pertains to the following: (B5) A method for preventing and/or alleviating and/or treating cytokine release syndrome and/or cytokine release, comprising administering a VEGF inhibitor to a subject. (B5-1) The method according to (B5), wherein the VEGF inhibitor is selected from an anti-VEGF antigen-binding molecule, an anti-VEGFR1 antigen-binding molecule, an anti-VEGFR2 antigen-binding molecule, a VEGF receptor, or a fusion protein comprising a fragment thereof, and one or more VEGF inhibitors of tyrosine kinase inhibitors. (B5-2) The method according to (B5) or (B5-1), wherein the VEGF inhibitor is one or more VEGF inhibitors selected from bevacizumab, ramucirumab, and aflibercept . (B5-3) The method according to any one of (B5) to (B5-2), wherein the cytokine release syndrome and/or cytokine release are cytokines accompanied by administration of an anti-T cell antigen-binding molecule Release syndrome and/or cytokine release. (B5-4) The method according to (B5-3), wherein the VEGF inhibitor is administered to the subject before, simultaneously with, or after the administration of the anti-T cell antigen-binding molecule. (B5-5) The method according to (B5-3), wherein the VEGF inhibitor is administered to the subject before or simultaneously with the administration of the anti-T cell antigen-binding molecule. (B5-6) The method according to any one of (B5-3) to (B5-5), 6 days before, 5 days before, 4 days before, 3 days before the administration of the anti-T cell antigen-binding molecule The aforementioned VEGF inhibitor is administered to the subject 1 day before, 2 days ago, 1 day before, or on the same day, before the administration of the aforementioned anti-T cell antigen-binding molecule. (B5-7) The method according to any one of (B5-3) to (B5-6), wherein the anti-T cell antigen-binding molecule is a bispecific antigen-binding molecule comprising: (1) a domain comprising an antibody variable region having T cell receptor complex binding activity, and (2) A domain comprising an antibody variable region having cancer antigen-binding activity. (B5-8) The method according to any one of (B5-3) to (B5-7), wherein the anti-T cell antigen-binding molecule is a bispecific antibody comprising: (1) a domain comprising an antibody variable region having T cell receptor complex binding activity, (2) a domain comprising an antibody variable region having glypican3-binding activity, and (3) A domain comprising an Fc region whose binding activity to an Fcγ receptor is reduced. (B5-9) The method according to any one of (B5) to (B5-8), for preventing and/or alleviating and/or treating cytokine release syndrome and/or cytokine release, and treating cancer . (B5-10) The method according to (B5-9), wherein the cytokine release syndrome is caused by the release of cytokines in either or both of the non-tumor tissue and the tumor tissue.

Alternatively, in a non-limiting aspect, the present disclosure relates to the following: (C1) A method of preventing and/or alleviating and/or treating cytokine release syndrome and/or cytokine release and treating cancer, comprising administering an anti-T cell antigen binding molecule and a VEGF inhibitor to a subject. (C1-1) A method of combined therapy for preventing and/or alleviating and/or treating cytokine release syndrome and/or cytokine release and treating cancer, comprising the step of administering an anti-T cell antigen-binding molecule to a subject , and the step of administering a VEGF inhibitor. (C1-2) The method according to (C1) or (C1-1), wherein the VEGF inhibitor is administered to the subject before, simultaneously with, or after the administration of the anti-T cell antigen-binding molecule. (C1-3) The method according to any one of (C1) to (C1-2), wherein the VEGF inhibitor is administered to the subject before or simultaneously with the administration of the anti-T cell antigen-binding molecule. (C1-4) The method according to any one of (C1) to (C1-3), wherein 6 days before, 5 days before, 4 days before the administration of the anti-T cell antigen-binding molecule to the subject, The aforementioned VEGF inhibitor was administered 3 days before, 2 days ago, 1 day before, or on the day before the administration of the aforementioned anti-T cell antigen-binding molecule. (C1-5) The method according to any one of (C1) to (C1-4), wherein the VEGF inhibitor is selected from anti-VEGF antigen-binding molecules, anti-VEGFR1 antigen-binding molecules, anti-VEGFR2 antigen-binding molecules, VEGF A receptor or a fusion protein comprising a fragment thereof, and one or more VEGF inhibitors of tyrosine kinase inhibitors. (C1-6) The method according to any one of (C1) to (C1-5), wherein the VEGF inhibitor is one or more selected from bevacizumab, ramucirumab, and aflibercept VEGF inhibitor. (C1-7) The method according to any one of (C1) to (C1-6), further comprising administering an immune checkpoint inhibitor to the subject. (C1-8) The method according to (C1-7), wherein the immune checkpoint inhibitor is an immune checkpoint inhibitor selected from an anti-PD1 antibody, an anti-PD-L1 antibody, and an anti-PD-L2 antibody. (C1-9) The method according to (C1-7) or (C1-8), wherein the immune checkpoint inhibitor is atezolizumab. (C1-10) The method according to any one of (C1) to (C1-9), wherein no corticosteroid is administered to the subject before or simultaneously with the administration of the anti-T cell antigen-binding molecule. (C1-11) The method according to any one of (C1) to (C1-9), wherein a corticosteroid is further administered to the subject before, simultaneously with or after the administration of the anti-T cell antigen-binding molecule. (C1-12) The method according to (C1-10) or (C1-11), wherein the corticosteroid is Dexamethasone, a pharmaceutically acceptable salt thereof, or a derivative thereof. (C1-13) The method according to any one of (C1) to (C1-12), wherein the anti-T cell antigen-binding molecule is a bispecific antigen-binding molecule comprising: (1) a domain comprising an antibody variable region having T cell receptor complex binding activity, and (2) A domain comprising an antibody variable region having cancer antigen-binding activity. (C1-14) The method according to any one of (C1) to (C1-13), wherein the anti-T cell antigen-binding molecule is a bispecific antibody comprising: (1) a domain comprising an antibody variable region having T cell receptor complex binding activity, (2) a domain comprising an antibody variable region having glypican3-binding activity, and (3) A domain comprising an Fc region whose binding activity to an Fcγ receptor is reduced. (C1-15) The method according to any one of (C1) to (C1-14), wherein the cytokine release syndrome and/or the cytokine release is a cell accompanied by the administration of the anti-T cell antigen-binding molecule Hormone release syndrome and/or cytokine release. (C1-16) The method according to any one of (C1) to (C1-15), wherein the cytokine release syndrome is caused by the release of cytokines in either or both of the non-tumor tissue and the tumor tissue.

In a non-limiting aspect, the present disclosure relates to a method comprising administering an anti-T cell antigen-binding molecule and a VEGF inhibitor for use in the various aspects of combination therapy disclosed herein, via the A method of treating cancer with concomitant therapy, and any one selected from the group consisting of prevention, alleviation, and treatment of cytokine release syndrome accompanied by administration of an anti-T cell antigen-binding molecule by the concomitant therapy, or its combination.

Or in a non-limiting aspect, the present disclosure is related to the following: (C2) A combination therapy for preventing and/or alleviating and/or treating cytokine release syndrome and/or cytokine release, and treating cancer, comprising an anti-T cell antigen binding molecule and a VEGF inhibitor. (C2-1) The combination therapy according to (C2), wherein the VEGF inhibitor is administered before, simultaneously with, or after the administration of the anti-T cell antigen-binding molecule. (C2-2) The combination therapy according to (C2) or (C2-1), wherein the VEGF inhibitor is administered before or simultaneously with the administration of the anti-T cell antigen-binding molecule. (C2-3) The combination therapy according to (C2) or (C2-1), 6 days before, 5 days before, 4 days before, 3 days before, and 2 days before the administration of the anti-T cell antigen-binding molecule The aforementioned VEGF inhibitor is administered before, 1 day before, or on the day before the administration of the aforementioned anti-T cell antigen-binding molecule. (C2-4) The combination therapy according to any one of (C2) to (C2-3), wherein the VEGF inhibitor is selected from an anti-VEGF antigen-binding molecule, an anti-VEGFR1 antigen-binding molecule, an anti-VEGFR2 antigen-binding molecule, VEGF receptor or a fusion protein comprising a fragment thereof, and one or more cytokine inhibitors of tyrosine kinase inhibitors. (C2-5) The combination therapy according to any one of (C2) to (C2-4), wherein the aforementioned VEGF inhibitor is one selected from bevacizumab, ramucirumab, and aflibercept The above VEGF inhibitors. (C2-6) The combination therapy according to any one of (C2) to (C2-5), further comprising using it together with an immune checkpoint inhibitor. (C2-7) The combination therapy according to (C2-6), wherein the immune checkpoint inhibitor is an immune checkpoint inhibitor selected from an anti-PD1 antibody, an anti-PD-L1 antibody, and an anti-PD-L2 antibody. (C2-8) The combination therapy according to (C2-6) or (C2-7), wherein the immune checkpoint inhibitor is atezolizumab. (C2-9) The combination therapy according to any one of (C2) to (C2-8), wherein no corticosteroid is administered before or simultaneously with the administration of the anti-T cell antigen-binding molecule. (C2-10) The combination therapy according to any one of (C2) to (C2-8), wherein a corticosteroid is further administered before, simultaneously with, or after the administration of the anti-T cell antigen-binding molecule. (C2-11) The combination therapy according to (C2-9) or (C2-10), wherein the corticosteroid is Dexamethasone, a pharmaceutically acceptable salt thereof, or a derivative thereof. (C2-12) The combination therapy according to any one of (C2) to (C2-11), wherein the anti-T cell antigen-binding molecule is a bispecific antigen-binding molecule comprising: (1) a domain comprising an antibody variable region having T cell receptor complex binding activity, and (2) A domain comprising an antibody variable region having cancer antigen-binding activity. (C2-13) The combination therapy according to any one of (C2) to (C2-12), wherein the anti-T cell antigen-binding molecule is a bispecific antibody comprising: (1) a domain comprising an antibody variable region having T cell receptor complex binding activity, (2) a domain comprising an antibody variable region having glypican3-binding activity, and, (3) A domain comprising an Fc region whose binding activity to an Fcγ receptor is reduced. (C2-14) The combination therapy according to any one of (C2) to (C2-13), wherein the cytokine release syndrome and/or cytokine release is accompanied by administration of the anti-T cell antigen-binding molecule Cytokine release syndrome and/or cytokine release. (C2-15) The combination therapy according to any one of (C2) to (C2-14), wherein the cytokine release syndrome is caused by the release of cytokines in either or both of the non-tumor tissue and the tumor tissue. (C3) A pharmaceutical composition for preventing and/or alleviating and/or treating cytokine release syndrome and/or cytokine release, and treating cancer, comprising an anti-T cell antigen binding molecule and a VEGF inhibitor. (C3-1) The pharmaceutical composition according to (C3), wherein the VEGF inhibitor is selected from an anti-VEGF antigen-binding molecule, an anti-VEGFR1 antigen-binding molecule, an anti-VEGFR2 antigen-binding molecule, a VEGF receptor, or a fusion comprising a fragment thereof A protein, and one or more cytokine inhibitors of tyrosine kinase inhibitors. (C3-2) The pharmaceutical composition according to (C3) or (C3-1), wherein the VEGF inhibitor is one or more VEGF selected from bevacizumab, ramucirumab, and aflibercept inhibitor. (C3-3) The pharmaceutical composition according to any one of (C3) to (C3-2), which is further used in combination with an immune checkpoint inhibitor. (C3-4) The pharmaceutical composition according to (C3-3), wherein the immune checkpoint inhibitor is an immune checkpoint inhibitor selected from an anti-PD1 antibody, an anti-PD-L1 antibody, and an anti-PD-L2 antibody. (C3-5) The pharmaceutical composition according to (C3-3) or (C3-4), wherein the immune checkpoint inhibitor is atezolizumab. (C3-6) The pharmaceutical composition according to any one of (C3) to (C3-5), wherein no corticosteroid is administered before or simultaneously with the administration of the pharmaceutical composition. (C3-7) The pharmaceutical composition according to any one of (C3) to (C3-5), wherein a corticosteroid is further administered before, simultaneously with, or after the administration of the pharmaceutical composition. (C3-8) The pharmaceutical composition according to (C3-6) or (C3-7), wherein the corticosteroid is Dexamethasone, a pharmaceutically acceptable salt thereof, or a derivative thereof. (C3-9) The pharmaceutical composition according to any one of (C3) to (C3-8), wherein the anti-T cell antigen-binding molecule is a bispecific antigen-binding molecule comprising: (1) a domain comprising an antibody variable region having T cell receptor complex binding activity, and (2) A domain comprising an antibody variable region having cancer antigen-binding activity. (C3-10) The pharmaceutical composition according to any one of (C3) to (C3-9), wherein the anti-T cell antigen-binding molecule is a bispecific antibody comprising: (1) a domain comprising an antibody variable region having T cell receptor complex binding activity, (2) a domain comprising an antibody variable region having glypican3-binding activity, and, (3) A domain comprising an Fc region whose binding activity to an Fcγ receptor is reduced. (C3-11) The pharmaceutical composition according to any one of (C3) to (C3-10), wherein the cytokine release syndrome and/or cytokine release is accompanied by administration of the anti-T cell antigen-binding molecule Cytokine Release Syndrome and/or Cytokine Release. (C3-12) The pharmaceutical composition according to any one of (C3) to (C3-11), wherein the cytokine release syndrome is caused by the release of cytokines in either or both of the non-tumor tissue and the tumor tissue . (C4) A combination of an anti-T cell antigen binding molecule and a VEGF inhibitor. [Effect of invention]

Anti-T cell antigen-binding molecules that bind to T cell antigens such as CD3 have attracted attention as a new cancer treatment method utilizing the antitumor effect of T cells possessed by living organisms. On the other hand, it has also been pointed out that by targeting cytokine-producing T cells, there is a possibility of causing side effects due to cytokines (eg, cytokine release syndrome; CRS). Therefore, there is concern that the administration of anti-T cell antigen-binding molecules may cause unexpected side effects due to cytokines. According to the present disclosure, because the anti-T cell antigen-binding molecule is administered in combination with a prior VEGF inhibitor, unexpected side effects due to cytokines can be prevented or reduced.

i. definition The following definitions are provided to facilitate understanding of the present invention described in this specification.

peculiarly "Specifically" refers to one molecule of molecules that specifically binds, and does not show any significant binding state with respect to molecules other than one or a plurality of binding partner molecules. Furthermore, it can also be used when a domain including an antibody variable region has specificity for a specific epitope among a plurality of epitopes contained in a certain antigen. Furthermore, when the epitope to which the domain including the antibody variable region binds is included in a plurality of different antigens, the antigen-binding molecule including the antibody variable region can bind to various antigens including the epitope. various antigens.

binding activity The term "binding activity" refers to the aggregate strength of non-covalent interactions between one or more binding sites of a molecule (eg, an antibody) and a binding partner (eg, an antigen) of the molecule . Here, "binding activity" is not strictly limited to a 1:1 interaction between members of a certain binding pair (eg, antibody and antigen). For example, this binding activity is specifically referred to as intrinsic binding affinity ("affinity") when reflecting the fact that the members of the binding pair interact 1:1 in a 1-valent manner. When the members of the binding pair are capable of both monovalent binding and multivalent binding, the binding activity is the sum of these binding forces. In general, the binding activity of a molecule X to its partner Y can be expressed as a dissociation constant (KD) or "the amount of analyte bound per unit amount of ligand" (hereinafter referred to as "binding amount"). Generally speaking, those with ordinary knowledge in the technical field to which the invention pertains should understand that the lower the value of the dissociation constant (KD), the higher the binding activity, and the higher the value of "the amount of analyte binding per unit amount of ligand" or "the amount of binding". The higher the binding activity, the higher the binding activity. The binding activity can be measured by a usual method known in the technical field including those described in the present specification. Specific examples and exemplified aspects for measuring binding activity (including affinity) will be described later. Specific examples and exemplified aspects for measuring binding affinity will be described later.

Anti-"target antigen"-binding antigen-binding molecule term "anti-"target antigen-binding antigen-binding molecule" (here, "target antigen" may be an antigen protein that can be targeted, for example, including T cell antigen, cancer antigen , cytokines, and cytokine receptors.), or "antigen-binding molecule that binds to a target antigen", refers to an antigen-binding molecule that is capable of binding to a target antigen with sufficient affinity, resulting in , the antigen-binding molecule is useful as a diagnostic and/or therapeutic agent when targeting the antigen. In one aspect, when the degree of binding of the antigen-binding molecule to an unrelated non-target antigenic protein is determined (eg, by radioimmunoassay (RIA)), the degree of binding of the antigen-binding molecule to the target antigen is less than that of the antigen-binding molecule. about 10%. In a specific aspect, the antigen-binding molecule that binds to the target antigen has ≦1 μM, ≦100 nM, ≦10 nM, ≦1 nM, ≦0.1 nM, ≦0.01 nM, or ≦0.001 nM (eg, 10 ⁻⁸ M or less, eg, 10 ^-8 M to 10 ^-13 M, eg, 10 ^-9 M to 10 ^-13 M) dissociation constant (Kd). In particular aspects, the antigen-binding molecule binds to an epitope of a target antigen retained between target antigens of different species.

Reference antigen-binding molecules and antigen-binding molecules that "bind to the same epitope" A reference antigen-binding molecule and an antigen-binding molecule that "binds to the same epitope" refer to an antigen-binding molecule that prevents more than 50% of the reference antigen-binding molecule from binding to its own antigen in a competition assay, or vice versa. Said that the reference antigen binding molecule prevented more than 50% of the binding of the antigen binding molecule to its own antigen in the competition assay. Exemplary competitive assays are provided in this specification.

In an exemplary competition assay, immobilized antigens (eg, GPC3, CD3, and/or VEGF) are included in a composition comprising a first labeled antigen-binding molecule that binds to the antigen, and a first labeled antigen-binding molecule for binding to the antigen. 1. The antigen-binding molecule was incubated in a solution of the second unlabeled antigen-binding molecule of the competition ability test. The second antigen-binding molecule was present in the supernatant of the fusion tumor. As a control, the immobilized antigen was incubated in a solution containing the first labeled antigen-binding molecule but not the second unlabeled antigen-binding molecule. After incubation under conditions that allow the first antigen-binding molecule to bind to the antigen, excess unbound antigen-binding molecule is removed, and the amount of the label bound to the immobilized antigen is measured. When the amount of the label bound to the immobilized antigen is substantially reduced compared to the control sample and the test sample, it is shown that the second antigen-binding molecule competes with the first antigen-binding molecule for binding to the antigen . See Harlow and Lane (1988) Antibodies: A Laboratory Manual ch. 14 (Cold Spring Harbor Laboratory, Cold Spring Harbor, NY).

antigen binding molecule The term "antigen-binding molecule" in this specification is used in the widest range, as long as it can exhibit the desired antigen-binding activity, including but not limited to, including monoclonal antibodies, polyclonal antibodies, and multispecific antibodies (for example, Bispecific antibodies), antibody derivatives and antibody fragments, various antibody constructs.

In a specific aspect, the antibodies provided herein can also be further modified to include additional non-proteinaceous moieties known in the art and readily available (this is referred to as an "antibody derivative"). Moieties suitable for derivatization of antibodies include, but are not limited to, water-soluble polymers. Non-limiting examples of water-soluble polymers include, but are not limited to, polyethylene glycol (PEG), ethylene glycol/propylene glycol copolymers, carboxymethyl cellulose, polydextrose (dextran), polyvinyl alcohol, poly Polyvinyl pyrrolidone, poly-1,3-dioxolane, poly 1,3,6-trioxane, ethylene/ Maleic anhydride copolymers, polyamino acids (either homopolymers or random copolymers), and polydextrose or poly(n-vinylpyrrolidone) polyethylene glycol, polypropylene glycol copolymers, Polypropylene oxide/ethylene oxide copolymers, polyoxyethylated polyols (eg, glycerol), polyvinyl alcohol, and mixtures of these. Polyethylene glycol propionaldehyde is advantageous for manufacture due to its stability in water. The polymer may have any molecular weight and may be branched or unbranched. The number of polymers attached to the antibody may also vary, and if more than one polymer is attached, these may be the same molecule or different molecules. In general, the number and/or type of polymers used for derivatization are not limited to these, but may depend on the specific properties or functions of the antibody to be improved, whether the antibody derivative is used in therapy under limited conditions, etc. Consider and decide.

Antibody Fragments An "antibody fragment" refers to a molecule other than the intact antibody comprising a portion of the intact antibody bound to the antigen to which the intact antibody binds. Examples of antibody fragments include, but are not limited to, Fv, Fab, Fab', Fab'-SH, F(ab') ₂ ; diabodies; linear antibodies; single chain antibody molecules (eg, scFv); and Multispecific antibodies formed from antibody fragments.

Reference antibodies and antibodies that bind to the same epitope Reference antibodies and antibodies that "bind to the same epitope" refer to antibodies that prevent more than 50% of the reference antibody's binding to its own antigen in a competition assay, or, conversely, a reference antibody that is in a competition assay Prevent more than 50% of the antibody from binding to its own antigen. Exemplary competitive assays are provided in this specification.

Chimeric antibody The term "chimeric" antibody means that a portion of the heavy and/or light chain is derived from a specific source or species, while the remainder of the heavy and/or light chain is derived from a different source or species. Antibody.

Antibody "class" The "class" of an antibody refers to the type of constant domain or constant region possessed by the heavy chain of the antibody. There are 5 major types of antibodies: IgA, IgD, IgE, IgG, and IgM. Also, some of them can be subdivided into subclasses (isotypes; homotypes). For example, IgG1, IgG2, IgG3, IgG4, IgA1, and IgA2. The heavy chain constant domains corresponding to different types of immunoglobulins are called α, δ, ε, γ, and μ, respectively.

The constant regions of each isotype of IgG1, IgG2, IgG3, and IgG4 are referred to as Cγ1, Cγ2, Cγ3, and Cγ4, respectively. The amino acid sequences of the polypeptides constituting the Fc regions of human Cγ1, Cγ2, Cγ3, and Cγ4 are shown in SEQ ID NOs: 23, 24, 25, and 26. The relationship between the amino acid residues constituting each amino acid sequence and the EU numbering of kabat (also referred to as EU INDEX in this specification) is shown in FIG. 1 .

Fc region In the present specification, the term "Fc region" is used to define the C-terminal region of an immunoglobulin heavy chain comprising at least a portion of the constant region. This term includes Fc regions of native-type sequences as well as variant Fc regions. In one embodiment, the human IgG heavy chain Fc region extends from Cys226 or Pro230 to the carboxy terminus of the heavy chain. However, the C-terminal lysine (Lys447) or glycine-lysine (Gly446-Lys447) of the Fc region may or may not be present. Unless otherwise limited in this specification, the numbering of amino acid residues in the Fc region or constant region is according to Kabat et al., Sequences of Proteins of Immunological Interest, 5th Ed. Public Health Service, National Institutes of Health, Bethesda, MD The EU numbering system (also known as the EU index) as described in 1991.

Fc receptors "Fc receptor" or "FcR" refers to a receptor that binds to the Fc region of an antibody. In some aspects, the FcR is a native human FcR. In some aspects, FcRs include those that bind to IgG antibodies (gamma receptors), receptors of the FcγRI, FcγRII, and FcγRIII subgroups, dual variants and alternative splicing of these receptors Shape. FcγRII receptors include FcγRIIA ("activating receptor") and FcγRIIB ("inhibiting receptor"), and mainly have different similar amino acid sequences in their cytoplasmic domains. The activating receptor FcγRIIA contains an immunoreceptor tyrosine-based activation motif (ITAM) in its cytoplasmic domain. The inhibitory receptor FcyRIIB contains an immunoreceptor tyrosine-based inhibition motif (ITIM) in its cytoplasmic domain (see, eg, Daeron, Annu. Rev. Immunol. 15:203-234 (1997). ) . An overview of FcRs is found, for example, in Ravetch and Kinet, Annu. Rev. Immunol 9:457-92 (1991); Capel et al., Immunomethods 4:25-34 (1994); and de Haas et al., J. Lab. Clin. Med 126:330-41 (1995). Other FcRs to be identified in the future are also included in the term "FcR" in this specification.

The term "Fc receptor" or "FcR" further includes the neonatal-type receptor FcRn, which is responsible for the movement of maternal IgG to the fetus (Guyer et al., J. Immunol. 117:587 (1976) and Kim et al., J. Immunol. 24:249 (1994)) and regulation of immunoglobulin homeostasis. Methods for measuring binding to FcRn are well known (for example, see Ghetie and Ward., Immunol. Today 18(12):592-598 (1997); Ghetie et al., Nature Biotechnology, 15(7):637-640 ( 1997); Hinton et al., J. Biol. Chem. 279(8):6213-6216 (2004); WO2004/92219 (Hinton et al.)).

In vivo binding to human FcRn and plasma half-life of human FcRn high-affinity binding polypeptides can be achieved by, for example, transfecting mice or transfected human cell lines with genes expressing human FcRn, Or administered to a primate with a polypeptide with a variant Fc region. WO2000/42072 (Presta) describes antibody variants with increased or decreased FcR binding. See also, eg, Shields et al. J. Biol. Chem. 9(2):6591-6604 (2001).

Fc region-containing antibody The term "Fc region-containing antibody" refers to an Fc region-containing antibody. The C-terminal lysine of the Fc region (residue 447 according to the EU numbering system) or the C-terminal glycine-lysine of the Fc region (residues 446-447) can be obtained via, for example, between purification of the antibody or encoding the antibody. removed by nucleic acid recombination. Thus, a composition comprising an antibody with an Fc region according to the present invention may comprise an antibody with G446-K447, an antibody with G446 but not K447, an antibody with G446-K447 completely removed, or a combination of the above three antibodies mixture.

The Fc region with reduced Fc receptor binding activity includes an Fc region with reduced binding activity to any Fcγ receptor of FcγI, FcγIIA, FcγIIB, FcγIIIA and/or FcγIIIB. The reduction of the binding activity to any Fcγ receptor of FcγI, FcγIIA, FcγIIB, FcγIIIA and/or FcγIIIB can also be determined by screening methods using ALPHA in addition to FACS and ELISA formats known to those skilled in the art to which the invention belongs. (Amplified Luminescent Proximity Homogeneous Assay), BIACORE method of surface plasmon resonance (SPR) phenomenon, etc. (Proc. Natl. Acad. Sci. USA (2006) 103 (11), 4005-4010).

An antigen-binding molecule or antibody comprising an Fc region with reduced binding activity to an Fc receptor includes an antigen-binding molecule or antibody with reduced effector function. Antigen-binding molecules or antibodies with reduced effector function comprise residues 238, 265, 269, 270, 297, 327, and 329 in the Fc region with one or more substitutions (US Pat. No. 6,737,056). Such Fc variants include so-called "DANA" Fc variants with alanine substitutions at residues 265 and 297 (US Pat. No. 7,332,581), including those with amino acid positions 265, 269, 270, 297, and 327 2 or more substituted Fc variants.

Specific antibody variants with increased or decreased binding to FcRs have been described. (See US Pat. No. 6,737,056; WO2004/056312, and Shields et al., J. Biol. Chem. 9(2): 6591-6604 (2001).)

In particular aspects, the antibody variant comprises one or more amino acid substitutions (eg, at positions 298, 333, and/or 334 (residues by EU numbering) of the Fc region) that concomitantly improve ADCC substitution) of the Fc region.

In some aspects, procedures in the Fc region result in altered (i.e., , either increased or decreased) changes in C1q binding and/or complement-dependent cellular damage (CDC).

In some aspects, the Fc region with reduced binding activity to an Fc receptor comprises an Fc variant of the Fc region exemplified in this specification.

framework "Framework" or "FR" refers to variable domain residues other than hypervariable region (HVR) residues. The FR of the variable domain is generally composed of four FR domains: FR1, FR2, FR3, and FR4. Accordingly, the sequences of HVR and FR generally appear in VH (or VL) in the following order: FR1-H1(L1)-FR2-H2(L2)-FR3-H3(L3)-FR4.

full-length antibody The terms "full-length antibody", "intact antibody", and "entire antibody" are used interchangeably in this specification and refer to an antibody having a structure substantially similar to that of a natural antibody, or having an Fc region as defined in this specification. heavy chain antibody.

host cell The terms "host cell", "host cell strain", and "host cell culture" are used interchangeably and refer to cells (including progeny of such cells) into which foreign nucleic acid has been introduced. Host cells include "transformants" and "transformed cells," which include the primary transformed cell and progeny derived from the cell regardless of the number of passages. The progeny also need not have the exact same nucleic acid content as the parental cell, and may also contain variations. Variant progeny that have the same function or biological activity as used at the time of screening or selection of the original transformed cells are also included in this specification.

human antibody A "human antibody" is an antibody produced by humans or human cells, or having amino acids corresponding to the amino acid sequence of an antibody from a non-human source using a human antibody repertory or other human antibody coding sequence sequence of antibodies. The definition of human antibody specifically excludes humanized antibodies comprising non-human antigen-binding residues.

Human Common Framework A "human consensus framework" is one that exhibits the most commonly occurring amino acid residues in a selected population of human immunoglobulin VL or VH framework sequences. Typically, human immunoglobulin VL or VH sequences are selected from a subclass of variable domain sequences. Subgroups of general sequences are those in Kabat et al., Sequences of Proteins of Immunological Interest, Fifth Edition, NIH Publication 91-3242, Bethesda MD (1991), vols. 1-3. In one aspect, for VL, the subgroup is subclass κI as described by Kabat et al., supra. In one aspect, for VH, the subgroup is subclass III as described by Kabat et al., supra.

humanized antibody "Humanized" antibodies include amino acid residues from non-human HVRs and amino acid residues from human FRs and are referred to as chimeric antibodies. In one aspect, the humanized antibody comprises substantially all of at least one, typically two, variable domains, in which all or substantially all of the HVRs (eg, CDRs) correspond to non-human antibodies , and all or substantially all of the FRs correspond to human antibodies. A humanized antibody may also optionally contain at least a portion of an antibody constant region from a human antibody. A "humanized form" of an antibody (eg, a non-human antibody) refers to an antibody that has been humanized.

hypervariable region The terms "hypervariable region", "HVR" or "CDR" as used in this specification refer to a sequence that is hypervariable ("complementarity determining region" or "complementarity determining region"), and/or forms a fixed structure loops ("hypervariable loops"), and/or regions of the variable domain of an antibody comprising antigen-contacting residues ("antigen-contacting"). Typically an antibody contains 6 CDRs: 3 for VH (H1, H2, H3), and 3 for VL (L1, L2, L3). The CDRs exemplified in this specification include the following: (a) at amino acid residues 26-32 (L1), 50-52 (L2), 91-96 (L3), 26-32 (H1), 53-55 (H2), and 96-101 (H3 ) generated at the hypervariable loop (Chothia and Lesk, J. Mol. Biol. 196:901-917 (1987)); (b) at amino acid residues 24-34 (L1), 50-56 (L2), 89-97 (L3), 31-35b (H1), 50-65 (H2), and 95-102 (H3 ) (Kabat et al., Sequences of Proteins of Immunological Interest, 5th Ed. Public Health Service, National Institutes of Health, Bethesda, MD (1991)); (c) at amino acid residues 27c-36 (L1), 46-55 (L2), 89-96 (L3), 30-35b (H1), 47-58 (H2), and 93-101 (H3 ) (MacCallum et al. J. Mol. Biol. 262: 732-745 (1996)); and, (d) contains amino acid residues 46-56 (L2), 47-56 (L2), 48-56 (L2), 49-56 (L2), 26-35 (H1), 26-35b (H1) , 49-65 (H2), 93-102 (H3), and 94-102 (H3), a combination of (a), (b), and/or (c). Unless otherwise indicated, CDR residues and other residues in the variable domain (eg, FR residues) are numbered in the present specification according to Kabat et al., supra.

individual An "individual", "subject" or "subject" is a mammal. Mammals include, but are not limited to, domesticated animals (such as cows, sheep, cats, dogs, horses), primates (such as humans, and non-human primates such as apes), rabbits, and rodents (such as mice and rat). In certain aspects, the individual or subject is a human.

isolated antigen binding molecule An "isolated" antigen-binding molecule refers to one that has been separated from components of its native environment. In some aspects, purification to More than 95% or 99% pure. For an overview of methods for assessing the purity of antibodies, see, eg, Flatman et al., J. Chromatogr. B 848:79-87 (2007).

isolated nucleic acid "Isolated" nucleic acid refers to a nucleic acid molecule that is separated from components of its original environment. The isolated nucleic acid includes a nucleic acid molecule contained in a cell that usually contains the nucleic acid molecule, and the nucleic acid molecule exists extrachromosomally or exists in a chromosomal location different from the original chromosomal location.

Isolated nucleic acid encoding antigen binding molecule "Isolated nucleic acid encoding an antigen-binding molecule" means one that encodes one or more polypeptide chains or fragments thereof (in the case of antibodies, heavy and light chains of antibodies or fragments thereof) of the antigen-binding molecule or plural nucleic acid molecules, including nucleic acid molecules carried on one vector or individual vectors, and nucleic acid molecules present in one or plural positions in a host cell.

monoclonal antibody The term "monoclonal antibody" used in the present specification refers to an antibody obtained from a substantially homogeneous antibody population. That is, the individual antibodies that make up the group, in addition to the mutant antibodies that can be generated (e.g., mutant antibodies that contain naturally occurring mutations, or mutant antibodies that occur in the manufacture of monoclonal antibody preparations, which usually exist several amounts), bind to the same and/or the same antigen-binding site. In contrast to polyclonal antibody preparations, which typically contain different antibodies to different determinants (epitopes), each monoclonal antibody of a monoclonal antibody preparation is a single determinant on an antigen. The adjective "monoclonal" is therefore characteristic of antibodies obtained from a substantially homogeneous population of antibodies and should not be construed as requiring any particular method for producing antibodies. For example, monoclonal antibodies used in accordance with the present invention, including but not limited to methods using transgenic animals comprising all or part of a fusion tumor method, recombinant DNA method, phage display method, human immunoglobulin loci, can be obtained through a variety of The preparation of the method, the method of preparing the monoclonal antibody, and other exemplary methods are described in the present specification.

accompanying documents The term "accompanying documents" is usually included in the commercial packaging of a therapeutic product for use in an instruction manual containing information on the indications, usage, dosage, method of administration, concomitant therapy, contraindications, and/or warnings for the use of this therapeutic product .

amino acid sequence identity The definition of "percent (%) amino acid sequence identity" for a reference polypeptide sequence is the percentage of amino acid residues in the reference polypeptide sequence and amino acid residues in the same candidate sequence, in order to obtain the maximum The percent sequence identity, alignment of the sequences and, if necessary, the insertion of spaces after the substitution, and any retained substitutions, are also not taken into account for sequence identity. Alignment for the purpose of determining percent amino acid sequence identity can use a variety of methods within the skill of the art, by using, for example, BLAST, BLAST-2, ALIGN, Megalign (DNA STAR) software or GENETYX (registered trademark) (GENETYX Co., Ltd.) and other publicly available computer software. Those skilled in the art to which the invention pertains can decide to include necessary algorithms and appropriate parameters for obtaining sequence alignment in order to achieve maximum alignment of the entire length of the aligned sequences.

The ALIGN-2 sequence alignment computer program is the work of Genentech Inc., and its source code and user documentation are submitted to the US Copyright Office (US Copyright Office, Washington D.C., 20559) at the same time, with the US Copyright Registration No. TXU510087 login. The ALIGN-2 program can be purchased publicly from Genentech, Inc., South San Francisco, California, or compiled from source code. ALIGN-2 programs are compiled for use on UNIX operating systems including Digital UNIX (UNIX is a registered trademark) V4.0D. All sequence alignment parameters were set by the ALIGN-2 program and were not changed. In the case of amino acid sequence alignment using ALIGN-2, the percent amino acid sequence identity of the administered amino acid sequence A that is identical to or aligned with the administered amino acid sequence B (or can also be explained An administered amino acid sequence A) that is identical to or aligned to an administered amino acid sequence B with or contains a certain percentage of amino acid sequence identity) is calculated via the formula: 100 of the fraction X/Y times. Here, X is the number of amino acid residues evaluated as identical by the sequence calibration program ALIGN-2, which is the same in the calibration of A and B, and Y is the number of amino acid residues in B all numbers. It should be understood that when the length of amino acid sequence A is not equal to the length of amino acid sequence B, A's percent amino acid sequence identity to B is the same as B's percent amino acid to A. Sequence identity is not the same. Unless otherwise specified, all percent amino acid sequence identity values used in the present disclosure are obtained using the ALIGN-2 computer program as described in the preceding paragraph.

pharmaceutical preparations The term "pharmaceutical preparation" refers to a preparation in a form that effectively exhibits the biological activity of the active ingredient contained therein, and does not contain additional requirements for toxicity that is unacceptable to the subject to which the preparation is administered. thing.

pharmaceutically acceptable carrier "Pharmaceutically acceptable carrier" means a component other than the active ingredient in a pharmaceutical preparation that is non-toxic to a subject. Pharmaceutically acceptable carriers include, but are not limited to, buffers, excipients, stabilizers, or preservatives.

treat As used in this specification, "treatment" (and its grammatical derivatives, such as "to treat", "to treat", etc.) refers to clinical interventions intended to alter the natural course of the individual being treated, and may also be used for the purpose of The prevention is carried out during the period when the clinical morbidity passes. Desired effects of treatment include, but are not limited to, prevention of disease occurrence or recurrence, alleviation of symptoms, reduction of any direct or indirect pathological effects due to disease, prevention of metastasis, slowing of disease progression, disease state recovery or alleviation, and relief or improved prognosis. In some aspects, the antibodies of the invention are used to delay the onset or delay the progression of a disease.

variable region The term "variable region" or "variable domain" refers to the domain of the heavy or light chain of an antibody that is involved in binding of an antibody to an antigen. The variable domains (VH and VL, respectively) of the heavy and light chains of native antibodies, typically each domain comprises 4 retained framework regions (FR) and 3 hypervariable regions (HVR), with similar structure. (For example, refer to Kindt et al., Kuby Immunology, 6th ed., W. H. Freeman and Co., page 91 (2007).) One VH or VL domain can sufficiently confer antigen-binding specificity. Furthermore, an antibody that binds to a specific antigen can also be isolated by screening the complementarity of the respective VL or VH domain from the VH or VL domain of the antibody that binds to the antigen. See, eg, Portolano et al., J. Immunol. 150: 880-887 (1993); Clarkson et al., Nature 352: 624-628 (1991).

vector The term "vector" used in the present specification refers to a nucleic acid molecule capable of adding another nucleic acid linked thereto. The term includes vectors that are self-replicating nucleic acid constructs, as well as vectors that are inserted into the genome of the host cell into which they are introduced. A vector is the nucleic acid itself capable of operatively expressing the linkage. Such vectors are referred to in this specification as "expression vectors".

multispecific antibody In certain aspects, the antigen-binding molecules (or antibodies) provided herein are multispecific antibodies (eg, bispecific antibodies). A multispecific antibody is a monoclonal antibody having at least two different binding specificities. Multispecific antibodies (eg, bispecific antibodies) can be prepared as full-length antibodies or antibody fragments.

Methods for making multispecific antibodies include, but are not limited to, recombinant co-expression of heavy chain-light chain pairs of two immunoglobulins with different specificities (cf. Milstein and Cuello, Nature 305:537 (1983). ), WO93/08829, and Traunecker et al., EMBO J. 10:3655 (1991)) and knob-in-hole techniques (see, eg, US Pat. No. 5,731,168). Multispecific antibodies can also be manipulated to produce electrostatic steering effects of Fc heterodimeric molecules (WO2009/089004A1); cross-linking two or more antibodies or fragments (see US Pat. No. 4,676,980 and Brennan et al., Science, 229:81 (1985)); production of antibodies with two specificities using a leucine zipper (see Kostelny et al., J. Immunol., 148(5):1547 -1553 (1992)); use "diabodies (DIABODY)" technology to produce bispecific antibody fragments (refer to Hollinger et al., Proc. Natl. Acad. Sci. USA, 90: 6444-6448 (1993)); and use of single-chain Fv (scFv) dimers (see Gruber et al., J. Immunol., 152: 5368 (1994)); and, for example, described in Tutt et al. J. Immunol. 147: 60 (1991) Trispecific antibodies are prepared for manufacture.

Modified antibodies with three or more functional antigen-binding sites including "octopus antibodies" are also included in the present specification (see US Patent Publication No. 2006/0025576 A1).

The antigen-binding molecule or fragment in the present specification includes one antigen-binding site that binds to another antigen different from the specific antigen, and also includes "dual acting Fab" or "DAF" (refer to U.S. Patent Publication No. 2008). /0069820).

II. Anti-T cell antigen binding molecule and pharmaceutical composition In one aspect, an "anti-T cell antigen-binding molecule" refers to an antigen-binding molecule that binds to an antigen on a T cell, including an antigen-binding molecule that binds to a T cell receptor complex. In one aspect, the anti-T cell antigen binding molecule is a multispecific antigen binding molecule. In one aspect, the anti-T cell antigen-binding molecule comprises "a domain comprising an antibody variable region having T cell receptor complex-binding activity" and "a domain comprising an antibody variable region having cancer antigen-binding activity" The bispecific antigen-binding molecule, preferably a bispecific antibody. In one aspect, the bispecific antibody may also have the configuration of a single chain antibody, eg, a configuration in which a linker is used to bind the variable region of the antibody. In one aspect, the anti-T cell antigen-binding molecule further comprises an Fc region with reduced binding activity to Fcγ receptors.

Furthermore, in one aspect, a domain comprising an antibody variable region having T cell receptor complex binding activity, preferably a domain comprising an antibody variable region having T cell receptor binding activity, more preferably comprising Domains of antibody variable regions with CD3 binding activity. Furthermore, in one aspect, the above-mentioned "domain comprising an antibody variable region" is provided by one or more variable domains of an antibody, and the domain comprising an antibody variable region preferably comprises an antibody light chain variable region (VL). ) and antibody heavy chain variable region (VH). As an example of such a domain comprising an antibody variable region, various antibody fragments, for example, "scFv (single chain Fv)", "single chain antibody (single chain antibody)", "Fv", " scFv2 (single chain Fv 2)", "Fab" or "F(ab')2", etc.

Domains comprising antibody variable regions with cancer antigen-binding activity In the present specification, "a domain comprising an antibody variable region having cancer antigen-binding activity" refers to a portion of an antibody comprising a complementary region that specifically binds to a part or all of a cancer antigen.

cancer specific antigen In the present specification, "cancer-specific antigen" means an antigen that can distinguish cancer cells from normal cells, and an antigen expressed by cancer cells, for example, an antigen expressed in association with malignant transformation of cells, including when cells become cancerous, Abnormal sugar chains appearing on the surface of cells and protein molecules. Specifically, for example, GPC3, ALK receptor (pleiotrophin receptor), pleiotrophin, KS 1/4 pancreatic cancer antigen, ovarian cancer antigen (CA125), prostatic acid phosphate, prostate specific antigen (PSA), melanin Tumor-associated antigen p97, melanoma antigen gp75, high molecular weight melanoma antigen (HMW-MAA), prostate specific membrane antigen, carcinoembryonic antigen (CEA), polymorphic epithelial mucin antigen, human milk fat globule antigen, CEA, Colorectal tumor-associated antigens such as TAG-72, CO17-1A, GICA 19-9, CTA-1 and LEA, Burkitt's lymphoma antigen-38.13, CD19, human B lymphoma antigen-CD20, CD33, ganglioside ( ganglioside) GD2, ganglioside GD3, ganglioside GM2 and ganglioside GM3 melanoma-specific antigen, tumor-specific transplantation cell surface antigen (TSTA), T antigen, DNA tumor virus and RNA tumor Virus-induced tumor antigens such as viral envelope antigens, colon CEA, 5T4 carcinoembryonic trophoblast glycoprotein, and carcinoembryonic antigen α-fetoprotein such as bladder tumor carcinoembryonic antigens, human lung cancer antigens Differentiation antigens such as L6 and L20, fibrosarcoma antigens, human leukemia T cell antigen-Gp37, neonatal glycoproteins, neurolipids, breast cancer antigens such as EGFR (epithelial growth factor receptor), NY-BR-16, Differentiation antigens such as NY-BR-16 and HER2 antigen (p185HER2), polymorphic epithelial mucin (PEM), malignant human lymphocyte antigen-APO-1, and I antigen found in fetal erythrocytes, in the early stage of adult erythrocyte discovery Endodermal I antigen, embryos before transplantation, I(Ma) found in gastric cancer, M18, M39 found in mammary epithelium, SSEA-1, VEP8, VEP9, Myl, VIM-D5 found in bone marrow cells, colorectal D156-22, TRA-1-85 (blood group H) found in cancer, SCP-1 found in testicular and ovarian cancer, C14 found in colon cancer, F3 found in lung cancer, AH6, Y half found in gastric cancer Antigen (hapten), Ley, TL5 (blood group A) found in embryonic cancer cells, EGF receptor found in A431 cells, E1 series (blood group B) found in pancreatic cancer, embryonic cancer cells FC10.2, gastric cancer antigen, CO-514 found in adenocarcinoma (blood group Lea), NS-10 found in adenocarcinoma, CO-43 (blood group Leb), G49 found in EGF receptor of A431 cells, MH2 (blood group ALeb/Ley) found in colon cancer, 19.9 found in colon cancer, gastric cancer mucin, T5A7 found in bone marrow cells, R24 found in melanoma, 4.2 found in embryonal carcinoma cells, GD3, D1.1, OFA-1, GM2, OFA-2, GD2, and M1:22:25:8 and SSEA-3 and SSEA-4 found in 4-8 cell stage embryos, subcutaneous T-cell lymphoma antigen, MART-1 antigen, Sialy Tn (STn) antigen, colon cancer Antigen NY-CO-45, lung cancer antigen NY-LU-12 variant A, adenocarcinoma antigen ART1, tumor-associated brain-testicular cancer antigen (cancer neuroantigen MA2, tumor-associated neuroantigen), nerve cancer ventral antigen 2 (NOVA2), blood cell cancer antigen gene 520, tumor-associated antigen CO-029, tumor-associated antigen MAGE-C1 (cancer/testis antigen CT7), MAGE-B1 (MAGE-XP antigen), MAGE-B2 (DAM6), MAGE-2, MAGE-4a, MAGE-4b, and MAGE-X2, cancer-testis antigen (NY-EOS-1), YKL-40, and fragments of any of the above-mentioned polypeptides or modified structures thereof, etc. (mentioned above modified phosphate groups, sugar chains, etc.), EpCAM, EREG, CA19-9, CA15-3, sialyl SSEA-1 (SLX), HER2, PSMA, CEA, CLEC12A, etc. The cancer-specific antigen that is the target of the cancer-specific antigen-binding domain of the present invention is particularly preferably one expressed on the cell surface. Examples of such cancer-specific antigen include CD19, CD20, EGFR, HER2, and EpCAM. , EREG.

Domains comprising antibody variable regions with GPC3 binding activity In the present specification, "a domain comprising an antibody variable region having Glypican3 (GPC3)-binding activity" means a portion of an antibody that specifically binds to a part or all of the above-mentioned GPC3 protein or a partial peptide thereof and is complementary to a domain .

In this specification, Glypican3 (GPC3) is a group of heparan sulfate proteoglycans bound to the cell surface via the polysaccharide phosphatidylinositol, that is, a protein belonging to the Glypican family (Filmus, J. Clin. Invest ., 2001, 108, 497-501). Glypican plays an important role in cell proliferation, differentiation and migration. GPC3 is expressed in more than 70% of hepatocellular carcinoma tissues obtained from surgical resection or biological specimens, but is completely absent, or almost absent, in adjacent nonneoplastic liver lesions and in most adult tissues (Zhu -Zu-W, Gut, 2001, 48, 558-564, Yamauchi, Mod. Pathol., 2005, 18, 1591-1598).

Domains comprising antibody variable regions with T cell receptor complex binding activity In the present specification, "a domain comprising an antibody variable region having T cell receptor complex binding activity" refers to a domain comprising a complementary domain that specifically binds to a part or all of the T cell receptor complex. Part of a cell receptor complex antibody. The T cell receptor complex may be the T cell receptor itself, or may be an adapter molecule that together with the T cell receptor constitutes the T cell receptor complex. As an adaptor molecule, CD3 is preferred.

Domains comprising antibody variable regions with T cell receptor binding activity In the present specification, "a domain comprising an antibody variable region having T cell receptor-binding activity" means a part of a T cell receptor antibody comprising a complementary region that specifically binds to a part or all of a T cell receptor . A part of the T cell receptor to which the domain of the present invention binds may be a variable region or a constant region, preferably an epitope present in the constant region. Examples of constant region sequences include T cell receptor alpha chain (SEQ ID NO: 9) of RefSeq accession number CAA26636.1, T cell receptor beta chain (SEQ ID NO: 10) of RefSeq accession number C25777, and RefSeq accession number 10. T cell receptor γ1 chain (SEQ ID NO: 11) of A26659, T cell receptor γ2 chain (SEQ ID NO: 12) of RefSeq accession number AAB63312.1, T cell receptor delta chain of RefSeq accession number AAA61033.1 (SEQ ID NO: 12) No.: 13) sequence.

Domains comprising antibody variable regions with CD3 binding activity In the present specification, "a domain comprising an antibody variable region having CD3-binding activity" means a part of a CD3 antibody that specifically binds to a part or all of CD3 and is complementary to the domain.

The domain comprising the variable region of an antibody having CD3-binding activity of the present invention may bind to any epitope if there is an epitope present in the sequence constituting the γ chain, δ chain, or ε chain of human CD3. By. In the present invention, it is preferable to use a light chain variable region (VL) of an anti-CD3 antibody and a heavy chain of an anti-CD3 antibody that bind to an epitope existing in the extracellular region of the epsilon chain of the human CD3 complex. Domains of the variable region (VH). For such domains, in addition to the light chain variable region (VL) of the anti-CD3 antibody and the heavy chain variable region (VH) of the anti-CD3 antibody described in the Examples, the OKT3 antibody (Proc. Natl. Acad) can also be preferably used. . Sci. USA (1980) 77, 4914-4917), or various CD3 binding domains comprising the light chain variable region (VL) of a CD3 antibody and the heavy chain variable region (VH) of a CD3 antibody. In addition, the γ chain, δ chain, or ε chain constituting human CD3 can be appropriately used to immunize a desired animal by the aforementioned method, and to obtain an antibody variable region derived from an anti-CD3 antibody having desired properties, which can be appropriately used. area. Anti-CD3 antibodies derived from domains comprising antibody variable regions having CD3-binding activity, as previously described, appropriately humanized or human antibodies can be used. The structure of the γ chain, δ chain, or ε chain constituting CD3, the polynucleotide sequences thereof are described in SEQ ID NO: 9 (NM_000073.2), 10 (NM_000732.4) and 11 (NM_000733.3), and the polypeptide sequences thereof are described In sequence numbers: 12 (NP_000064.1), 13 (NP_000723.1) and 14 (NP_000724.1) (RefSeq accession numbers are shown in parentheses).

The domain including the antibody variable region in the antigen-binding molecule of the present invention can bind to the same epitope. Here, the same epitope can exist in the protein formed by the amino acid sequence described in SEQ ID NO: 2 or SEQ ID NO: 14. Alternatively, the domains comprising the variable regions of antibodies in the antigen-binding molecules of the present invention can bind to different epitopes. Here, different epitopes can exist in the protein formed by the amino acid sequence described in SEQ ID NO: 2 or SEQ ID NO: 14.

Fc region with reduced binding activity to Fcγ receptors In one aspect, the reduction in the binding activity to Fcγ receptors refers to, for example, that the binding activity of the test antigen-binding molecule is 50% or less, preferably 45% or less, compared with the binding activity of the control antigen-binding molecule. , 40% or less, 35% or less, 30% or less, 20% or less, 15% or less, preferably 10% or less, 9% or less, 8% or less, 7% or less, 6% or less, 5% or less, 4% less than 3%, less than 2%, less than 1% of binding activity.

As a control antigen-binding molecule, an antigen-binding molecule having an Fc region of an IgG1, IgG2, IgG3, or IgG4 monoclonal antibody can be appropriately used. Furthermore, when an antigen-binding molecule having a variant of the Fc region of an antibody of a specific isotype is used as a test substance, it is possible to use an antigen-binding molecule having the Fc region of an antibody of the specific isotype as a control. The effect on the binding activity of the Fcγ receptor due to the variation possessed by the variant was examined. As described above, an antigen-binding molecule having a variant of an Fc region whose binding activity to an Fcγ receptor has been examined can be reduced can be appropriately prepared.

As examples of such variants, deletions of 231A-238S of specific amino acids according to EU numbering are known (WO 2009/011941); C226S, C229S, P238S, (C220S) (J. Rheumatol (2007) 34, 11); C226S, C229S (Hum. Antibod. Hybridomas (1990) 1 (1), 47-54); variants of C226S, C229S, E233P, L234V, L235A (Blood (2007) 109, 1185-1192).

That is, among the amino acids constituting the Fc region of an antibody of a specific isotype, preferably, amino acids having any of the following specified by EU numbering can be listed; bits, 233 bits, 234 bits, 235 bits, 236 bits, 237 bits, 238 bits, 239 bits, 240 bits, 264 bits, 265 bits, 266 bits, 267 bits, 269 bits, 270 bits, 295 bits, 296 bits, Antigen-binding molecules with Fc regions substituted at positions 297, 298, 299, 300, 325, 327, 328, 329, 330, 331, and 332. The isotype of the antibody derived from the Fc region is not particularly limited, and an Fc region derived from an IgG1, IgG2, IgG3, or IgG4 monoclonal antibody can be appropriately used, and an Fc region derived from an IgG1 antibody is preferably used.

For example, among the amino acids constituting the Fc region of an IgG1 antibody, any of the following substitutions specified by the EU numbering can be appropriately used (the numbers indicate the positions of the amino acid residues specified by the EU numbering, respectively). , the one-letter amino acid symbol before the number represents the amino acid residue before substitution, the one-letter amino acid symbol after the number represents the Fc region of the substituted amino acid residue), or at position 231 An antigen-binding molecule with a deleted Fc region in the amino acid sequence to position 238; (a) L234F, L235E, P331S, (b) C226S, C229S, P238S, (c) C226S, C229S, (d) C226S, C229S, E233P, L234V, L235A (e) L234A, L235A or L235R, N297A (f) L235A or L235R, S239K, N297A.

Furthermore, among the amino acids constituting the Fc region of an IgG2 antibody, it can be suitably used with any one of the following substitutions specified by EU numbering (the numbers indicate the positions of amino acid residues specified by EU numbering, respectively). , the single-letter amino acid symbol before the number represents the amino acid residue before substitution, and the single-letter amino acid symbol after the number represents the substituted amino acid residue) of the antigen-binding molecule of the Fc region; (g) H268Q, V309L, A330S, P331S (h)V234A (i) G237A (j) V234A, G237A (k)A235E, G237A (l) V234A, A235E, G237A.

In addition, among amino acids constituting the Fc region of an IgG3 antibody, it can be appropriately used with any one of the following substitutions specified by EU numbering (the numbers represent the amino acid residues specified by EU numbering, respectively). position, the one-letter amino acid symbol before the number represents the amino acid residue before substitution, and the one-letter amino acid symbol after the number represents the substituted amino acid residue) of the antigen-binding molecule of the Fc region; (m)F241A (n)D265A (o) V264A.

Furthermore, among amino acids constituting the Fc region of an IgG4 antibody, it can be appropriately used with any one of the following substitutions specified by EU numbering (the numbers represent the amino acid residues specified by EU numbering, respectively). position, the one-letter amino acid symbol before the number represents the amino acid residue before substitution, and the one-letter amino acid symbol after the number represents the substituted amino acid residue) of the antigen-binding molecule of the Fc region; (p)L235A, G237A, E318A (q)L235E (r) F234A, L235A.

As another preferred example, among the amino acids constituting the Fc region of the IgG1 antibody, there can be mentioned any one of the following amino acids specified by EU numbering: 233-position, 234-position, 235-position, 236-position, 237-position An antigen-binding molecule in which the Fc region of the amino acid corresponding to the EU numbering of the corresponding IgG2 or IgG4 is substituted at positions 327, 330, and 331.

Other preferred examples include amino acids having any one or more of the following specified by EU numbering among amino acids constituting the Fc region of an IgG1 antibody: 234th, 235th, 1st At position 297, the antigen-binding molecule of the Fc region substituted with other amino acids. The type of amino acid present after substitution is not particularly limited, but an antigen-binding molecule having an Fc region in which any one or more amino acids at positions 234, 235, and 297 are substituted with alanine is particularly preferred.

As another preferred example, among the amino acids constituting the Fc region of IgG1 antibodies, amino acids having any one of the following specified by EU numbering can be preferably cited: Fc substituted with another amino acid at the 265th position region of antigen-binding molecules. The type of amino acid present after substitution is not particularly limited, but an antigen-binding molecule having an Fc region in which the amino acid at position 265 is substituted with alanine is particularly preferred.

Bispecific antibody against Glypican3 (GPC3)/T cell receptor complex In one aspect, the anti-T cell antigen-binding molecule comprises (1) a domain comprising an antibody variable region having Glypican3 binding activity, (2) a domain comprising an antibody variable region having T cell receptor complex binding activity , and (3) a bispecific antibody comprising a domain of an Fc region whose binding activity to an Fcγ receptor is reduced. In one aspect, the antibody L chain variable region included in the antibody variable region having Glypican3-binding activity and the antibody variable region having T-cell receptor complex-binding activity of the present invention is preferably obtained to have the ability to bind to Glypican3. An L chain common to the binding energies of both the H chain having binding activity to the T cell receptor complex and the H chain having binding activity to the T cell receptor complex was used as the common L chain variable region of the bispecific antibody.

Examples of the antibody H chain variable region included in the antibody variable region having the preferred Glypican3-binding activity in the present disclosure include, for example, the antibody H chain variable regions described in Table 1, or the antibody H chain variable regions having CDR1, CDR2, and CDR3. An antibody H chain variable region having the same amino acid sequence as the CDR1, CDR2 and CDR3 amino acid sequences of the antibody H chain variable region described in Table 1, or an antibody H chain variable region having the same function as the variable region Antibody H chain variable region.

[Table 1]

Furthermore, as a preferred antibody variable region having a T cell receptor complex binding activity in the present disclosure, an antibody variable region having a binding activity to a T cell receptor can be exemplified. Among the T cell receptors, CD3 is preferred, and CD3ε is particularly preferred. Examples of the antibody H chain variable region included in such an antibody variable region include the antibody H chain variable regions described in Table 2, or the amino acid sequences of CDR1, CDR2, and CDR3 possessed by the antibody H chain variable regions described in Table 2, for example. An antibody H chain variable region having a CDR sequence identical to the amino acid sequence of CDR1, CDR2, and CDR3, or an antibody H chain variable region having the same function as the variable region.

[Table 2]

The relationship between the CDR regions of the amino acid residues constituting the amino acid sequence of the antibody H chain and the Kabat numbering is shown in FIG. 2 .

Examples of the common L chain variable region used in the present invention include the L chain variable regions described in Table 3, or the amino acid sequences of CDR1, CDR2, and CDR3 possessed by the antibody L chain described in Table 3. An antibody L chain variable region having CDR1, CDR2, and CDR3 amino acid sequences that are identical in amino acid sequences to an antibody L chain variable region, or an antibody L variable region having the same function as the variable region.

[table 3]

The relationship between the CDR regions of the amino acid residues constituting the amino acid sequence of the antibody L chain and the Kabat numbering is shown in FIG. 4 .

In a preferred aspect, as a combination of an antibody variable region having Glypican3 binding activity and an antibody variable region having T cell receptor complex binding activity, for example, the antibody H chain variable regions described in Table 4 can be listed. A combination of regions, or an antibody H chain having a CDR1, CDR2, and CDR3 amino acid sequence identical to the CDR1, CDR2, and CDR3 amino acid sequences of the antibody H chain variable regions described in Table 4 can be used. A combination of variable regions, or a combination of antibody H chain variable regions with the same function as the variable region.

[Table 4]

For the combination of the above-mentioned antibody variable region having Glypican3-binding activity and antibody variable region having T-cell receptor complex-binding activity, examples of preferred common L chains include L0000, L0011, L0201, L0203, and L0204. , L0206, L0208, L0209, L0211, L0212, L0222, or CDRs having the same amino acid sequence of CDR1, CDR2 and CDR3 of the common L chain as those of CDR1, CDR2 and CDR3 of the common L chain The common L chain of the sequence. As a specific preferred combination, for example, the combination of the antibody H chain variable region and the common L chain described in Table 5, or the combination of the amino acid sequence of CDR1, CDR2, and CDR3 with the antibody described in Table 5 can be mentioned. Combination of antibody variable regions having CDR1, CDR2, and CDR3 amino acid sequences that are identical to the amino acid sequences of variable regions and common L chains, or antibody H chain variable regions and common L chains that have the same function as the variable region The combination.

[table 5]

The Fc region contained in the anti-T cell antigen-binding molecule (preferably a bispecific antibody) is not particularly limited as long as its binding activity to Fcγ receptors is reduced, and preferred Fc regions include, for example, The combination of the Fc region part of E22Hh and the Fc region part of E22Hk, the combination of the Fc region part of E2702GsKsc and the Fc region part of E2704sEpsc, the combination of the Fc region part of E2702sKsc and the Fc region part of E2704sEpsc.

ERY974 In one aspect, the anti-T cell antigen binding molecule of the present disclosure is ERY974. ERY974 is a domain comprising (1) a domain comprising an antibody variable region having Glypican3 binding activity, (2) a domain comprising an antibody variable region having T cell receptor complex binding activity, and (3) a domain comprising Fcγ receptor A bispecific antibody that binds to an Fc region with reduced activity, comprising TR01H113 (SEQ ID NO: 168) as the heavy chain variable region on the CD3 side, E2702sKsc (SEQ ID NO: 62) as the heavy chain constant region on the CD3 side, GCH065 (SEQ ID NO: 206) as the heavy chain variable region on the GPC3 side, E2704sEpsc (SEQ ID NO: 61) as the heavy chain constant region on the GPC3 side, L0011 (SEQ ID NO: 223) as the common light chain variable region , and k0 (SEQ ID NO: 63) as a common light chain constant region. In other aspects, ERY974 respectively comprises the amino acid sequence of SEQ ID NO: 402 for the CD3 side chain, the amino acid sequence of SEQ ID NO: 385 for the GPC3 side chain, and the amino acid sequence of SEQ ID NO: 410 for the common light chain. .

In one aspect, amino acids included in the amino acid sequences of the present disclosure may be post-translationally modified. For example, the N-terminal glutamic acid residue (Q) is subjected to pyroglutamination to pyroglutamic acid (pGlu), a modification well known to those skilled in the art. Of course, the case where such an amino acid is modified after translation is also included in the amino acid sequence described in the present disclosure.

In the present disclosure, examples of other preferable anti-T cell antigen-binding molecules include bispecific antibodies comprising an antibody variable region having binding activity to Glypican3 and an antibody variable region having binding activity to CD3ε. More preferably, compared with the bispecific antibody GPC3_ERY22_rCE115 (disclosed in Example 1 of WO2015156268), the cytotoxic activity is the same or more preferably. Examples of such a bispecific antibody include a bispecific antibody having the H chain and L chain described in Example 3 (Table 13) of WO2015156268, or an antigen that binds to a repeating epitope bound to the antibody. The epitope is a bispecific antibody having an Fc region whose binding activity to Fcγ receptors is reduced as described above.

Here, in the present disclosure, "functionally the same" means the same binding affinity to an antigen or, when used as a bispecific antibody, the same cytotoxic activity against cells expressing Glypican3 or tissues containing the cells. The binding affinity and cytotoxic activity can be measured based on the description of the present specification. A desired cell expressing GPC3 or a desired tissue containing the cell can be used as the cell for measuring the cytotoxic activity, for example, PC-10 or NCI-H446, a human cancer cell line expressing GPC3, can be used. The same applies to the decrease in the binding activity to the Fcγ receptor in the antibody constant region.

For example, an antibody H chain variable region having the same function as the antibody H chain variable region (original H chain variable region) described in the specification of the present application refers to a pair of the original H chain with the variable region described in the specification of the present application. When the antibody L chain variable regions are combined, the binding affinity is the same, or when used as a bispecific antibody, the cytotoxic activity against Glypican3-expressing cells or tissues containing the cells is the same. In addition, the antibody L chain variable region having the same function as the antibody L chain variable region (original L chain variable region) described in the specification of the present application refers to the pair of the original L chain as described in the specification of the present application. When combined with the variable regions of the antibody H chain, the binding affinity is the same, or when used as a bispecific antibody, the cytotoxic activity against Glypican3-expressing cells or tissues containing the cells is the same.

Furthermore, "equivalent" does not necessarily require the same degree of activity, and can also enhance activity. Specifically, in the case of the binding affinity of the antigen, the value (KD value/parental KD value) compared with the binding affinity (parental KD value) of the antibody variable region of the control can be 1.5 or less. Condition. The value of KD value/parental KD value is preferably 1.3 or less, more preferably 1.2 or less, 1.1 or less, 1.0 or less, 0.9 or less, 0.8 or less, 0.7 or less, 0.6 or less, or 0.5 or less. The lower limit is not limited, for example, it can be ^10-1 , ^10-2 , ^10-3 , ^10-4 , ^10-5 , or ^10-6 . Specifically, in the present invention, the value of KD value/parental KD value is preferably 10 ^-6 to 1.5×10 ^-0 , more preferably 10 ^-6 to 10 ^-1 , more preferably 10 ^-6 to 10 ^-2 , more preferably 10 ^-6 ~10 ^-3 . In the case of cytotoxic activity, the value (cell growth inhibition rate/parental cell growth inhibition rate) compared with the cell growth inhibition rate of the control bispecific antibody (parent cell growth inhibition rate) can be cited as 0.7 the above situation. The concentration of the added multispecific antigen-binding molecule can be appropriately determined, and it is preferably measured, for example, at 0.01 nM, 0.05 nM, 0.1 nM, 0.5 nM, or 1 nM, preferably 0.05 nM or 0.1 nM. The value of cell growth inhibition rate/parental cell growth inhibition rate is preferably 0.8 or more, more preferably 0.9 or more, 1.0 or more, 1.2 or more, 1.5 or more, 2 or more, 3 or more, 5 or more, 10 or more, or 20 or more . The upper limit is not limited, such as 10, 10 ² , 10 ³ , 10 ⁴ , 10 ⁵ , or 10 ⁶ .

Furthermore, in the case of cytotoxic activity, the value (cell 50% growth inhibitory concentration/ When the parental cell 50% growth inhibitory concentration) is 1.5 or less. The 50% growth inhibitory concentration refers to the concentration of the multispecific antigen-binding molecule required to halve the cell proliferation rate compared to the case where the multispecific antigen-binding molecule is not added. The value of "50% cell growth inhibitory concentration/50% cell growth inhibitory concentration of parent" is preferably 1.3 or less, more preferably 1.2 or less, 1.1 or less, 1.0 or less, 0.9 or less, 0.8 or less, 0.7 or less, 0.6 or less, or less than 0.5. The lower limit is not limited, and may be, for example, 10 ^-1 , 10 ^-2 , 10 ^-3 , 10 ^-4 , 10 ^-5 , or 10 ^-6 . Specifically, it is preferably 10 ^-6 ~1.5x10 ^-0 , more preferably 10 ^-6 ~10 ^-1 , more preferably 10 ^-6 ~10 ^-2 , more preferably 10 ^-6 ~10 ^-3 .

Furthermore, regarding the domain comprising the antibody variable region having GPC3-binding activity, the KD value of GPC3 (for example, human GPC3) may be, for example, 5× ^{10 −9} M or less, preferably 4×10 ⁻⁹ M or less, for example, 3×10 ⁻⁹ M M or less, 2x10 ^-9 M or less, 1x10 ^-9 M or less, 8x10 ^-10 M or less, 5x10 ^-10 M or less, 4x10 ^-10 M or less, 3x10 ^-10 M or less, 2x10 ^-10 M or less, 1x10 ^-10 M or less , 8x10 ^-11 M or less, 5x10 ^-11 M or less, 4x10 ^-11 M or less, 3x10 ^-11 M or less, 2x10 ^-11 M or less, 1x10 ^-11 M or less, 8x10 ^-12 M or less, 5x10 ^-12 M or less, 4x10 ^-12 M or less, 3x10 ^-12 M or less, 2x10 ^-12 M or less, 1x10 ^-12 M or less, 8x10 ^-13 M or less, 5x10 ^-13 M or less, 4x10 ^-13 M or less, 3x10 ^-13 M or less, 2x10 ^-13 M or less, or 1x10 ^-13 M or less.

Furthermore, with regard to a domain comprising an antibody variable region having T cell receptor complex binding activity, for a human T cell receptor complex, such as a human T cell receptor, more specifically, a KD value such as a human CD3ε chain, For example, it can be 2x10 ^-7 M or less, preferably 1.5x10 ^-7 M or less, such as 1.4x10 ^-7 M or less, 1.3x10 ^-7 M or less, 1.2x10 ^-7 M or less, 1x10 ^-7 M or less, 3x10 ^-8 M or less, 2x10 ^-8 M or less, 1x10 ^-8 M or less, 8x10 ^-9 M or less, 5x10 ^-9 M or less, 4x10 ^-9 M or less, 3x10 ^-9 M or less, 2x10 ^-9 M or less, 1x10 ^-9 M or less , 8x10 ^-10 M or less, 5x10 ^-10 M or less, 4x10 ^-10 M or less, 3x10 ^-10 M or less, 2x10 ^-10 M or less, 1x10 ^-10 M or less, 8x10 ^-11 M or less, 5x10 ^-11 M or less, 4x10 ^-11 M or less, 3x10 ^-11 M or less, 2x10 ^-11 M or less, 1x10 ^-11 M or less, 8x10 ^-12 M or less, 5x10 ^-12 M or less, 4x10 ^-12 M or less, 3x10 ^-12 M or less, 2x10 ^-12 M or less, or 1x10 ^-12 M or less.

The bispecific antibodies of the present disclosure preferably have KD values for human GPC3 and human T cell receptor complexes (eg, human CD3ε chain) of ^5x10-9 M or less and ^2x10-7 M or less, respectively, more preferably 1x10 ^-9 M below and 5x10 ^-8 M below.

In the present invention, the "functionally identical" antibody variable region refers to the antibody H chain variable region and/or the antibody L chain variable region that satisfy the above-mentioned conditions, and is not particularly limited. As such antibody variable regions, for example, in the amino acid sequences of the variable regions described in Tables 1 to 3 above, one or a plurality of amino acids (for example, 1, 2, 3, 4, 5, or 10) amino acid) substitution, deletion, addition and/or insertion. In the amino acid sequence, a method known to those skilled in the art for substituting, deleting, adding, and/or inserting one or a plurality of amino acids is a method of introducing mutation into a protein. For example, one skilled in the art can use site-directed mutagenesis (Hashimoto-Gotoh, T, Mizuno, T, Ogasahara, Y, and Nakagawa, M. (1995) An oligodeoxyribonucleotide-directed dual amber method for site-directed mutagenesis. Gene 152 , 271-275, Zoller, MJ, and Smith, M. (1983) Oligonucleotide-directed mutagenesis of DNA fragments cloned into M13 vectors. Methods Enzymol. 100, 468-500, Kramer, W, Drutsa, V, Jansen, HW, Kramer, B, Pflugfelder, M, and Fritz, HJ (1984) The gapped duplex DNA approach to oligonucleotide-directed mutation construction. Nucleic Acids Res. 12, 9441-9456, Kramer W, and Fritz HJ (1987) Oligonucleotide-directed construction of mutations via gapped duplex DNA Methods. Enzymol. 154, 350-367, Kunkel, TA(1985) Rapid and efficient site-specific mutagenesis without phenotypic selection. Proc Natl Acad Sci U S A. 82, 488-492) et al. A variable region having the same function as an antibody variable region having the above-mentioned functions can be prepared by introducing appropriate variation in the base acid sequence.

In the case of altering the amino acid sequence, it is desirable to mutate to other amino acids that retain the nature of the amino acid side chain. For example, the properties of the amino acid side chain can include, for example, hydrophobic amino acids (A, I, L, M, F, P, W, Y, V), hydrophilic amino acids (R, D, N, C, E, Q, G, H, K, S, T), amino acids with aliphatic side chains (G, A, V, L, I, P), amino acids with hydroxyl-containing side chains (S, T, Y), amino acids (C, M) with side chains containing sulfur atoms, amino acids (D, N, E, Q) with side chains containing carboxylic acids and amides, An amino acid (R, K, H) containing a basic side chain, and an amino acid (H, F, Y, W) containing an aromatic side chain (any one in parentheses represents the single letter mark). Substitutions of amino acids within these groups are referred to as retained substitutions. Polypeptides with amino acid sequences modified by deletion, addition and/or substitution of other amino acid residues to one or more amino acid residues of an amino acid sequence are known to maintain their biological activity (Mark, D. F. et al., Proc. Natl. Acad. Sci. USA (1984) 81:5662-6; Zoller, M. J. and Smith, M., Nucleic Acids Res. (1982) 10:6487-500; Wang, A. et al., Science (1984) 224:1431-3; Dalbadie-McFarland, G. et al., Proc. Natl. Acad. Sci. USA (1982) 79:6409-13). The variable region of the present invention comprising such amino acid variation has at least 70%, more preferably 75%, of the CDR sequence, FR sequence or the amino acid sequence of the entire variable region of the variable region before the variation, More preferably at least 80%, more preferably at least 85%, even more preferably at least 90%, most preferably at least 95% amino acid sequence identity. The identity of the sequences in this specification is defined as the amine groups of the original H chain variable region or L chain variable region after introducing an appropriate gap (gap) and arranging the sequences as necessary to maximize the sequence identity. The residues of the acid sequence are the same residue ratio. The identity of the amino acid sequence can be determined by the method described later.

In addition, the "functionally identical antibody variable region" can also be obtained from, for example, a nucleic acid consisting of a nucleotide sequence encoding the amino acid sequence of the variable region described in Tables 1 to 3, or a nucleic acid that hybridizes under stringent conditions. get. For isolating nucleic acid hybridized under stringent conditions from a nucleic acid formed by a base sequence encoding an amino acid sequence of a variable region, stringent hybridization conditions may be, for example, 6M urea, 0.4% SDS, 0.5 × SSC, 37°C or the same stringent hybridization conditions. More stringent conditions, such as using 6M urea, 0.4% SDS, 0.1 × SSC, and 42°C, can be expected to isolate nucleic acids with higher similarity. Washing conditions after hybridization, such as 0.5×SSC (1×SSC is 0.15M NaCl, 0.015M sodium citrate, pH7.0), and washing at 0.1% SDS, 60°C, preferably 0.2×SSC, and Wash at 0.1% SDS, 60°C, preferably 0.2×SSC, and wash at 0.1% SDS, 62°C, preferably 0.2×SSC, and wash at 0.1% SDS, 65°C, preferably 0.1 ×SSC, and washed with 0.1% SDS at 65°C. Identification of the sequence of the isolated nucleic acid can be performed by a known method described later. The identity of the isolated nucleic acid is at least 50% or more, more preferably 70% or more, more preferably 90% or more (eg, 95%, 96%, 97%, 98%, 99% or more) of the entire base sequence. sex. In addition to the method using the above hybridization technique, a gene amplification method using primers synthesized based on base sequence information encoding the amino acid sequence of the variable region, such as the polymerase chain reaction (PCR) method, can be used to separate and A nucleic acid that hybridizes under stringent conditions with a nucleic acid formed from a base sequence encoding an amino acid sequence of a variable region.

The identity of the base sequence and the amino acid sequence can be determined according to Karlin and Altschul's algorithm BLAST (Proc. Natl. Acad. Sci. USA (1993) 90:5873-7). Based on this algorithm, programs called BLASTN, BLASTX were developed (Altschul et al., J. Mol. Biol. (1990) 215:403-10). In the case of analyzing the base sequence based on BLAST and BLASTN, the parameters are set to, for example, score=100 and wordlength=12. Also, in the case of analyzing the amino acid sequence by BLASTX based on BLAST, the parameters are set to, for example, score=50 and wordlength=3. In the case of BLAST and Gapped BLAST programs, the default parameters for each program are used. A specific method of such an analysis method is known (see the website of BLAST (Basic Local Alignment Search Tool) of NCBI (National Center for Biotechnology Information); http://www.ncbi.nlm.nih.gov).

Pharmaceutical composition containing anti-T cell antigen-binding molecule In another aspect, the present disclosure provides an anti-T cell antigen-binding molecule, preferably comprising (1) a domain comprising an antibody variable region having Glypican3 binding activity, (2) a domain comprising a T cell receptor complex binding activity A pharmaceutical composition comprising an antibody variable region domain of , and (3) a bispecific antibody comprising an Fc region domain with reduced Fcγ receptor-binding activity as active ingredients. Furthermore, the present disclosure relates to a cytotoxic pharmaceutical composition containing the anti-T cell antigen-binding molecule as an active ingredient. The pharmaceutical composition of the present disclosure induces such cell damage, especially T cell-dependent cell damage, and this activity is preferably administered to individuals who are preventing or suffering from a disease in need of treatment, or individuals with a possibility of recurrence.

As previously described specifically, human glypican3 (GPC3) is a cancer antigen that is specifically expressed in cancer cells. Therefore, a pharmaceutical composition comprising, as an active ingredient, an anti-T cell antigen-binding molecule comprising a domain of an antibody variable region having glypican3-binding activity, is preferably used for treatment, and prevention of either or both of them. That is, the present disclosure provides a pharmaceutical composition comprising the following bispecific antibody as an active ingredient, comprising: (1) a domain comprising an antibody variable region having glypican3-binding activity, ( 2) A domain comprising an antibody variable region having T cell receptor complex binding activity, and (3) a domain comprising an Fc region having reduced Fcγ receptor binding activity, for co-administration with a VEGF inhibitor A pharmaceutical composition for either or both of the treatment and prevention of cancer, a pharmaceutical composition selected from either or a combination thereof for the purpose of prevention, mitigation and treatment, the prevention, mitigation, and treatment To prevent, alleviate, and treat either or both of CRS and cytokine release resulting from the administration of the aforementioned pharmaceutical compositions.

Furthermore, the present disclosure relates to the manufacture of a pharmaceutical composition comprising (1) a domain comprising an antibody variable region having glypican3-binding activity, and (2) an antibody variable having T-cell receptor complex-binding activity. Co-administration of a bispecific antibody comprising an Fc region having a reduced binding activity to an Fcγ receptor and a VEGF inhibitor for either treatment or prevention of cancer, or The pharmaceutical compositions of both parties are used to select any one or a combination thereof for the purpose of prevention, alleviation, and treatment, and the prevention, alleviation, and treatment are caused by the administration of the aforementioned pharmaceutical compositions. Either or both of CRS and cytokine release are administered. Alternatively, the present disclosure relates to a method comprising (1) a domain comprising an antibody variable region having glypican3 binding activity, (2) a domain comprising an antibody variable region having T cell receptor complex binding activity, and (3) A method of co-administering a bispecific antibody comprising a domain of an Fc region with reduced Fcγ receptor-binding activity and a VEGF inhibitor for use in either or both of cancer treatment and prevention, for selection From any one of, or a combination of, the purpose of prevention, mitigation, and treatment, the prevention, mitigation, and treatment is the prevention, mitigation, and treatment of CRS and cytokine release resulting from the administration of the aforementioned pharmaceutical composition. either or both.

Further, the present disclosure provides a use comprising (1) a domain comprising an antibody variable region having glypican3 binding activity, (2) a domain comprising an antibody variable region having T cell receptor complex binding activity, and (3) ) Use of a bispecific antibody comprising a domain of an Fc region whose binding activity to Fcγ receptors is reduced, in either or both of the treatment and prevention of cancer co-administered with a VEGF inhibitor for use in selected from any one, or a combination thereof, for the purpose of preventing, alleviating, and treating CRS and cytokine release resulting from the administration of the aforementioned bispecific antibodies either or both.

Therefore, in a preferred aspect of the present disclosure, a step of identifying a subject suffering from GPC3-expressing cancer can be included before administration of the pharmaceutical composition. Cancer tissue is collected from a living body, cancer cells contained in the collected tissue are identified, and the expression level of GPC3 is determined by a known method. According to needs, it can be judged that the expression level of GPC3 is high when the expression level of GPC3 in the cells used as the sample is higher than the expression level of GPC3 in normal tissues. A preferable normal tissue for comparison of the expression level is a normal cell of the same tissue (organ) as the tissue (organ) from which the cancer cells are derived. More specifically, another aspect of the present disclosure provides a method for either or both of the treatment and prevention of cancer, comprising the step of judging a subject suffering from a cancer with high GPC3 expression, and the step of co-administering a bispecific antibody comprising 1) a domain comprising an antibody variable region having glypican3 binding activity, and (2) a domain comprising a T cell receptor An antibody variable region domain having a complex binding activity, and (3) a domain comprising an Fc region having reduced Fcγ receptor binding activity, selected from any one of the purpose of prevention, mitigation, and treatment, or a combination thereof, the prevention, alleviation, and treatment is the prevention, alleviation, and treatment of either or both of CRS and cytokine release resulting from administration of the aforementioned bispecific antibodies.

Furthermore, in the present disclosure, including: (1) a domain comprising an antibody variable region having Glypican3-binding activity, (2) a domain comprising an antibody variable region having T cell receptor complex binding activity, and, (3) A cytotoxicity inducer and a cell growth inhibitor containing a multispecific antigen-binding molecule comprising a domain of an Fc region whose binding activity to Fcγ receptors is reduced as an active ingredient can also be expressed as containing the anti-T cell antigen-binding agent. A method of inducing cytotoxicity in the step of administering the molecule to an individual, or the use of the anti-T cell antigen-binding molecule in the manufacture of a cytotoxicity-inducing agent and a cell proliferation inhibitor.

In the present disclosure, "contains (1) a domain comprising an antibody variable region having Glypican3 binding activity, (2) a domain comprising an antibody variable region having T cell receptor complex binding activity, and (3) a domain comprising "A multispecific antigen-binding molecule in the Fc region with reduced Fcγ receptor-binding activity as an active ingredient" means that the anti-T cell antigen-binding molecule is included as a main active ingredient, and does not limit the anti-T cell antigen The content of binding molecules.

Furthermore, the anti-T cell antigen-binding molecules of the present invention, preferably bispecific antibodies, can be encapsulated in microcapsules (microcapsules of hydroxymethyl cellulose, gelatin, poly[methyl methacrylate], etc.) as needed, as Colloidal drug delivery systems (liposomes, albumin microspheres, microemulsions, nanoparticles and nanocapsules, etc.) (refer to "Remington's Pharmaceutical Science ^16th edition", Oslo Ed. (1980) etc.). Furthermore, a method of using a drug as a sustained-release drug is also known, and this method can be applied to the anti-T cell antigen-binding molecule of the present invention (J. Biomed. Mater. Res. (1981) 15, 267-277, Chemtech. (1982) 12, 98-105, US Patent No. 3773719, European Patent Publication Nos. EP58481 and EP133988, Biopolymers (1983) 22, 547-556).

The pharmaceutical composition comprising the anti-T cell antigen-binding molecule of the present disclosure can be administered to a patient via either oral or parenteral administration. Preferably it is administered parenterally. Specific examples of the administration method include injection administration, nasal administration, transpulmonary administration, and percutaneous administration. Examples of the injection administration include intravenous injection, intramuscular injection, intraperitoneal injection, subcutaneous injection, and the like. For example, by injection, the pharmaceutical composition of the present invention, or the cell injury inducer and cell proliferation inhibitor can be administered systemically or locally. In addition, an appropriate administration method can be selected according to the patient's age and symptoms. The administration amount can be selected, for example, within the range of 0.0001 mg to 1000 mg per 1 kg of body weight per administration. Alternatively, for example, an administration amount in the range of 0.001 mg/body to 100,000 mg/body per patient may be selected. However, the pharmaceutical composition of the present invention, or the cell injury inducer and cell proliferation inhibitor are not limited to these doses.

The pharmaceutical composition comprising the anti-T cell antigen-binding molecule of the present disclosure can be formulated according to conventional methods (eg, Remington's Pharmaceutical Science, latest edition, Mark Publishing Company, Easton, U.S.A.), and may also contain pharmaceutically acceptable carriers or additives together. thing. For example, surfactants, excipients, colorants, fragrances, preservatives, stabilizers, buffers, suspending agents, isotonic agents, binding agents, disintegrating agents, lubricants, flow enhancers, flavoring agents, and the like. Furthermore, it is not limited to these, Other common carriers can be used suitably. Specifically, light anhydrous silicic acid, lactose, crystalline cellulose, mannitol, starch, calcium carboxymethyl cellulose, sodium carboxymethyl cellulose, hydroxypropyl cellulose, and hydroxypropyl methyl cellulose can be mentioned. , polyvinyl acetal diethyl aminoacetate, polyvinyl pyrrolidone, gelatin, medium chain fatty acid triglycerides, polyoxyethylene hardened castor oil 60, sugar, carboxymethyl cellulose, corn Powder, inorganic salts, etc. as carriers.

In one aspect, the present disclosure pertains to pharmaceutical compositions comprising anti-T cell antigen binding molecules, such as bispecific antibodies, Preferably, a bispecific antibody that binds to a cancer antigen and CD3, for use in a pharmaceutical composition for combined therapy with a VEGF inhibitor, the prevention, alleviation, and treatment are caused by the aforementioned bispecific Either or both of CRS and cytokine release by the administration of a sexual antibody. In one aspect, the VEGF inhibitor is, for example, capable of being selected from an anti-VEGF antigen-binding molecule, an anti-VEGFR1 antigen-binding molecule, an anti-VEGFR2 antigen-binding molecule, a VEGF receptor or a fusion protein comprising a fragment thereof, and a tyrosine kinase inhibitor formed groups. Among them, antibodies that bind to VEGF, or VEGFR1, VEGFR2, etc. are preferred VEGF inhibitors. Specifically, Bevacizumab is a humanized anti-VEGF antibody, or Ramucirumab is an anti-VEGFR2 human antibody. Other molecular-targeted drugs developed by targeting VEGF, such as Aflibercept, are also preferred examples of VEGF inhibitors. Details of the combination therapy are described in "V. Combination therapy" in this specification.

III. VEGF inhibitor In one aspect, "VEGF inhibitor" or "VEGF antagonist" refers to a substance that blocks, inactivates, or reduces the expression or level of activity of VEGF. A VEGF inhibitor is, for example, a compound, eg, an antagonist, that binds to VEGF, partially or fully masks, reduces, prevents, delays activation, inactivation, desensitization, or downregulates its activity or expression. In other examples, the VEGF inhibitor is a receptor that binds to VEGF, or blocks the binding of VEGF to a receptor, thereby partially or fully masking, reducing, preventing, delaying activation, inactivation, Compounds that desensitize, or downregulate, their activity or performance. VEGF inhibitors include antigen-binding molecules, antibodies, antibody derivatives, antibody fragments, polypeptide inhibitors of soluble receptors, etc., derivatives of these, and nucleic acid inhibitors of siRNA or antisense RNA, etc. Derivatives, genetically modified forms of soluble factors, such as forms having altered activities, naturally occurring and synthetic soluble factor antagonists, low molecular weight compounds, etc., but not limited to these.

In one aspect, the VEGF inhibitor is selected from one of an anti-VEGF antigen-binding molecule, an anti-VEGFR1 antigen-binding molecule, an anti-VEGFR2 antigen-binding molecule, a VEGF receptor or a fusion protein comprising a fragment thereof, and a tyrosine kinase inhibitor The above VEGF inhibitors. In one aspect, the VEGF inhibitor is one selected from an anti-VEGF antigen-binding molecule, an anti-VEGFR1 antigen-binding molecule, an anti-VEGFR2 antigen-binding molecule, a VEGF receptor or a fusion protein comprising a fragment thereof, or a tyrosine kinase inhibitor The above signal transduction factors or antibodies, antibody derivatives or antibody fragments that bind to their receptors.

VEGF signaling inhibitor In one aspect, the VEGF inhibitor is an inhibitor of VEGF signaling, eg, an inhibitor of VEGF or a VEGF receptor. In one aspect, the inhibitor can also be an anti-VEGF antigen binding molecule, anti-VEGF antibody (including chimeric anti-VEGF antibody, humanized anti-VEGF antibody, human anti-VEGF antibody), antigen-binding fragment or antibody derivative thereof. In yet another aspect, the inhibitor is an anti-VEGF receptor (VEGFR) antigen-binding molecule, an anti-VEGFR antibody (including a chimeric anti-VEGFR antibody, a humanized anti-VEGFR antibody, a human anti-VEGFR antibody) or an antigen-binding fragment thereof, or Antibody Derivatives. In one aspect, these inhibitors may also be soluble VEGF receptors or fragments thereof capable of blocking VEGF signaling. In another embodiment, the VEGF receptor or fragment thereof can also be a heterodomain, eg, a fusion protein fused to an Fc domain, eg, a VEGFR-Fc fusion protein.

In one aspect, the anti-VEGF antibody comprises, eg, Bevacizumab, Ranibizumab, and the like. In yet another aspect, the anti-VEGF receptor antibody comprises, eg, Ramucirumab. In one aspect, the inhibitor of VEGF signaling comprises a recombinant fusion protein of Aflibercept and the like.

Bevacizumab Bevacizumab is a humanized anti-VEGF antibody. Bevacizumab is an antibody represented by Japanese Accepted Names for Pharmaceuticals (JAN) Bevacizumab, CAS Registry No. 216974-75-3, or INN Name Bevacizumab. Bevacizumab is also a genetically recombinant humanized monoclonal antibody, which is composed of a complementarity-determining portion of a mouse anti-human vascular endothelial growth factor (VEGF) monoclonal antibody, a human framework portion, and a human IgG1 constant portion. Bevacizumab was produced by Chinese hamster ovary cells. Bevacizumab is composed of 2 H chains (γ1 chains) composed of 453 amino acid residues and 2 L chains (κ chains) composed of 214 amino acid residues Glycoprotein (molecular weight: about 149,000). "Bevacizumab" also includes the biosimilar bevacizumab.

Pharmaceutical composition containing VEGF inhibitor From another viewpoint, the present disclosure provides a pharmaceutical composition containing a VEGF inhibitor as an active ingredient, preferably an antibody against VEGF or its receptor, more preferably bevacizumab or ramucirumab . In another aspect, the present disclosure provides a pharmaceutical composition comprising a fusion protein comprising the extracellular domain of a VEGF receptor as an active ingredient, preferably aflibercept. In addition, the present disclosure relates to a pharmaceutical composition containing the VEGF inhibitor as an active ingredient for use in a pharmaceutical composition selected from any one or a combination thereof for the purpose of prevention, mitigation, and treatment. , and treatment is the prevention, alleviation, and treatment of either or both symptoms of cytokine release syndrome (CRS) and cytokine release. The pharmaceutical composition of the present disclosure is preferably administered to an individual who is likely to develop CRS or release of cytokines, and/or develop CRS or symptoms of CRS in need of treatment. In one aspect of the present disclosure, the mechanism that brings about CRS, cytokine release (or impending release) in a subject is arbitrary. That is, for whatever reason, the VEGF inhibitor can be administered to a subject for whom CRS, cytokine release is observed, or predicted to occur. For example, when there is a possibility of CRS or cytokine release due to the administration of a certain drug, a subject to which the drug has been administered (or to which it has been administered) can be selected, and the VEGF inhibitor can be administered. For example, for a subject who has observed CRS and cytokine release due to the administration of a certain drug in the past, when the same drug is administered again, there is a possibility that CRS and cytokine release may occur.

In one aspect of the present disclosure, there is provided a step of selecting subjects who have observed CRS or cytokine release due to the administration of a certain drug in the past, and who are scheduled to be administered the same drug or who have already been administered, and steps for selecting the subject for which the same drug has been administered. A subject, a VEGF inhibitor co-administered with the aforementioned agent. Alternatively, the present disclosure provides the use of a VEGF inhibitor co-administered with the aforementioned drug in order to administer the same drug to a subject who has observed CRS or cytokine release due to administration of a drug in the past. Furthermore, the present disclosure provides a method for selecting for the purpose of preventing, alleviating, and treating any one or a combination of the following symptoms selected from CRS and cells caused by the administration of the same agent. At least one symptom of low hormone release, comprising the steps of selecting a subject who has observed CRS and cytokine release due to the administration of a certain drug in the past, and the subject of administering the same drug, and co-administering the subject with the aforementioned drug VEGF inhibitor steps. According to the present disclosure, among subjects, measures that may lead to CRS and cytokine release include administration of anti-T cell antigen-binding molecules, such as bispecific antibodies, preferably to cancer antigens and CD3, to the subject. Bispecific antibodies.

Not intended to be limiting, the pharmaceutical composition of the present disclosure containing a VEGF inhibitor as an active ingredient can be used as a mechanism for the following purpose of inhibiting the release of IL-6 by inhibiting VEGF signaling, and the above purpose is selected from Either or a combination of preventing or reducing and treating CRS, cytokine release, or both. Without limitation, the pharmaceutical composition containing a VEGF inhibitor as an active ingredient in the present disclosure can be used as another mechanism for the purpose of inhibiting vascular permeability through a VEGF inhibitor and inhibiting the production of cytokines. Penetration from blood vessels into tissues, the above-mentioned purpose is selected from any one or a combination of preventing or alleviating and treating CRS.

Therefore, a pharmaceutical composition containing a VEGF inhibitor as an active ingredient in the present disclosure includes the step of administering the VEGF inhibitor to an individual, for selecting any one or a combination thereof for the purpose of prevention, alleviation, and treatment. Methods for preventing, alleviating, and treating either or both of CRS and cytokine release concomitant with administration of anti-T cell antigen binding molecules, and, for manufacture to select from Use of the VEGF inhibitor for any one or a combination of medicaments for the purpose of prevention, mitigation, and treatment concomitant with administration of an anti-T cell antigen binding molecule either or both of CRS and cytokine release.

In the present disclosure, "containing a VEGF inhibitor as an active ingredient" means that the VEGF inhibitor is contained as a main active ingredient, and is not intended to limit the content rate of the VEGF inhibitor.

Furthermore, the VEGF inhibitor of the present invention can be encapsulated in microcapsules (microcapsules of hydroxymethyl cellulose, gelatin, poly[methyl methacrylate], etc.) as needed, as a colloidal drug delivery system (liposome, albumin, etc.) microspheres, microemulsions, nanoparticles and nanocapsules, etc.) (refer to "Remington's Pharmaceutical Science ^16th edition", Oslo Ed. (1980) etc.). Furthermore, a method of using a drug as a sustained-release drug is also known, and this method can be applied to the VEGF inhibitor of the present invention (J. Biomed. Mater. Res. (1981) 15, 267-277, Chemtech. (1982) 12, 98-105, US Patent No. 3773719, European Patent Publication Nos. EP58481 and EP133988, Biopolymers (1983) 22, 547-556).

The pharmaceutical composition comprising the VEGF inhibitor of the present disclosure can be administered to a patient via either oral or parenteral administration. Parenteral administration is preferred. Specific examples of the administration method include injection administration, nasal administration, transpulmonary administration, and percutaneous administration. Examples of the injection administration include intravenous injection, intramuscular injection, intraperitoneal injection, subcutaneous injection, and the like. For example, by injection, the pharmaceutical composition of the present invention, or the cell injury inducer and cell proliferation inhibitor can be administered systemically or locally. In addition, an appropriate administration method can be selected according to the patient's age and symptoms. The administration amount can be selected, for example, within the range of 0.0001 mg to 1000 mg per 1 kg of body weight per administration. Alternatively, for example, an administration amount in the range of 0.001 mg/body to 100,000 mg/body per patient can be selected. However, the pharmaceutical composition of the present invention is not limited to these dosages.

In one aspect, when the VEGF inhibitor is bevacizumab, the active ingredient is about 5 to 15 mg/kg each time. For example, for patients with a body weight of 30 kg or more, 5 mg/kg or less can be selected each time. amount, 7.5 mg/kg or less, 10 mg/kg or less, 15 mg/kg or less, for example, intravenously over 0.5 hours, 1 hour, 2 hours, or 3 hours Administered by intramuscular injection. In one aspect, when the VEGF inhibitor is ramucirumab, the active ingredient is about 4 to 15 mg/kg each time, for example, for a patient with a body weight of 30 Kg or more, 8 mg/kg or The amounts below, 10 mg/kg or below, are administered via intravenous injection over, for example, 0.5 hour, 1 hour, 2 hours, or 3 hours. In different aspects, bevacizumab and ramucirumab are administered for the purpose of CRS treatment, and the symptoms of CRS after the first administration of bevacizumab and ramucirumab to patients If there is no improvement, up to 3 additional doses of bevacizumab and ramucirumab can be administered. In one example, when bevacizumab or ramucirumab is continuously administered, an interval of 8 hours or more is required from the previous administration.

The pharmaceutical composition comprising the VEGF inhibitor of the present disclosure can be formulated according to conventional methods (eg, Remington's Pharmaceutical Science, latest edition, Mark Publishing Company, Easton, U.S.A.), and may also contain pharmaceutically acceptable carriers or additives together. For example, surfactants, excipients, colorants, fragrances, preservatives, stabilizers, buffers, suspending agents, isotonic agents, binding agents, disintegrating agents, lubricants, flow enhancers, flavoring agents, and the like can be exemplified. Furthermore, it is not limited to these, Other common carriers can be used suitably. Specifically, light anhydrous silicic acid, lactose, crystalline cellulose, mannitol, starch, calcium carboxymethyl cellulose, sodium carboxymethyl cellulose, hydroxypropyl cellulose, and hydroxypropyl methyl cellulose can be mentioned. , polyvinyl acetal diethyl aminoacetate, polyvinyl pyrrolidone, gelatin, medium chain fatty acid triglycerides, polyoxyethylene hardened castor oil 60, sugar, carboxymethyl cellulose, corn Powder, inorganic salts, etc. as carriers.

In one aspect, the present disclosure relates to a pharmaceutical composition for use in combination therapy with an anti-T cell antigen binding molecule, comprising a VEGF inhibitor, preferably a VEGF inhibitor, more preferably directed against VEGF, its receptor. The best antibodies are bevacizumab and ramucirumab. Furthermore, the present disclosure relates to a pharmaceutical composition for use in combination therapy with an anti-T cell antigen-binding molecule, comprising a VEGF inhibitor composed of a fusion protein containing a ligand-binding site of a VEGF receptor, preferably a Bercept as the active ingredient. In one aspect, the anti-T cell antigen binding molecule is, for example, a bispecific antibody, preferably a bispecific antibody that binds to a cancer antigen and CD3. The details of the combination therapy are described in "V. Combination therapy" in this specification.

IV. Cytokine Release Syndrome (CRS) In one aspect, cytokine release syndrome (CRS) is administered with an agent such as an antibody drug (eg, anti-T cell antibody) or T cell drug (eg, chimeric antigen receptor (CAR) Severe and life-threatening side effects of T cells (CAR-T cells). Through the administration of antibody pharmaceuticals, T cell pharmaceuticals, etc., the immune response in the body is activated more than necessary, and inflammatory cytokines are released, which cause chills, nausea, fatigue, headache, fever, tachycardia, and blood pressure. Symptoms such as changes. In severe cases, there is a condition called a cytokine storm. CRS is the result of the activation of a high amount of immune cells that release inflammatory cytokines when many lymphocytes and/or myeloid cells are activated. The severity of CRS, that is, the onset of symptoms, varies depending on the level of immune cell activation in an individual, the type of drug administered, and/or the amount of systemic tumor tissue. In the case of T cell therapy for cancer, the onset of symptoms, such as in the case of a peak in T cell proliferation in vivo, is typically between days and weeks after administration of the T cell therapy. See, for example, Lee et al. Blood. 124.2 (2014): 188-95.

In one aspect, symptoms of CRS also include neurotoxicity, disseminated intravascular coagulation dysfunction, cardiac dysfunction, adult respiratory distress syndrome, renal failure, and/or liver failure. For example, symptoms of CRS may also include: fever, high fever with or without chills, fatigue, malaise, muscle pain, vomiting, headache, nausea, loss of appetite, joint pain, diarrhea, rash, hypoxia, excessive Respiration, hypotension, enlarged pulse pressure, potential decrease in stroke volume (late phase), increase in stroke volume (prephase), hyperazotemia, hypofibrinogenemia with or without hemorrhage, D-dimer rise, hyperbilirubinemia, hypertransaminemia, confusion, delirium, changes in mental status, hallucinations, tremors (shakes), convulsions, changes in pace, difficulty recalling single words, marked aphasia, or Dementia.

In one aspect, CRS is characterized by elevated concentrations of certain cytokines in an individual. In one aspect, the cytokine comprises IL-1β, IL-2, IL-4, IL-5, IL-6, IL-8, IL-10, IL-12, IL-13, IL-15, IL -17, IL-1Ra, IL-2R, IFN-α, IFN-γ, MIP-1α, MIP-1β, MCP-1, TNFα, GM-CSF, G-CSF, CXCL9, CXCL10, CXCR factor, VEGF, RANTES, eosin, EGF, HGF, FGF-β, CD30, CD30L, CD40, CD40L, ferritin, RAGE, but not limited thereto. Cytokines preferably comprise IL-6, IL-1 beta, or TNF-alpha, or any combination of these. The cytokine is more preferably IL-6. In one aspect, patients with larger tumor volumes have a higher incidence and severity of cytokine syndrome.

In one aspect, those skilled in the art should understand the term "cytokine concentration" to include the measured value of concentration, the size of the fold change, the size of the percentage (%) change, or the size of the change ratio. Furthermore, methods for measuring cytokines in blood, saliva, serum, urine, plasma, and/or serum are well known in the art.

Severity of CRS (Grade) In one aspect, CRS can be classified as severe (Grade) from 1 to 5. In one aspect, only symptomatic treatment (eg, nausea, fever, fatigue, muscle pain, malaise, headache) is required for CRS at Grade 1, and symptoms are not life-threatening. With regard to grade 2 CRS, symptoms require, and in general, respond to, modest intervention. Individuals with grade 2 CRS develop hypotensive symptoms responsive to fluids or 1 low-dose blood pressure-raising agent; or they develop grade 2 epigastric toxicity or to low flow oxygen (less than 40% oxygen) Responsive mild respiratory symptoms. In individuals with grade 3 CRS, hypotension is generally not reversible with fluid therapy or 1 low-dose blood pressure raising agent. Such individuals generally require lower flow rates of oxygen, have grade 3 organ toxicity (eg, renal or cardiac dysfunction or blood coagulation disturbance) and/or grade 4 hypertransaminemia. Individuals with grade 3 CRS require more aggressive interventions, such as requiring greater than 40% oxygen, high doses of blood pressure-lowering agents, and/or multiple blood pressure-lowering agents. Individuals with grade 4 CRS contain grade 4 organ toxicity or the need for mechanical artificial ventilation, and suffer from symptoms that are immediately life-threatening. Individuals with grade 4 CRS generally do not have hypertransaminemia. In individuals with grade 5 CRS, toxicity was the cause of death.

In one aspect, CRS grades are given based on the Common Terminology Criteria for Adverse Events (CTCAE) v4.03 shown in Table 6 and the Common Terminology of Adverse Events shown in Table 7. Benchmark v5.0 (Common Terminology Criteria for Adverse Events (CTCAE) v5.0), Lee's Criteria in 2014 shown in Table 8 (Lee DW, et al. Current concepts in the diagnosis and management of cytokine release syndrome. Blood , 124 (2014), pp. 188-195), the 2019 ASTCT CRS Consensus Grading shown in Table 9 (Lee DW, et al. ASTCT Consensus Grading for Cytokine Release Syndrome and Neurologic Toxicity Associated with Immune Effector Cells. Biol Blood Marrow Transplant. 2019 Apr;25(4):625-638), Penn's guidelines, MSKCC's guidelines, CARTOX's guidelines (above Biol Blood Marrow Transplant. 2019 Apr;25(4):625-638) and proceed. The assignment of a grade, preferably CRS, is based on CTCAE v 4.03, CTCAE v 5.0, or Lee's 2014 guidelines. Unless otherwise specified, the CRS used in this specification refers to the CRS in accordance with the criteria in Table 8 (Lee's criteria in 2014.

[Table 6] (CTCAE v 4.03) Grade I Mild reaction; no need to interrupt the infusion; no treatment required Grade II Treatment or interruption of the drip is required. But responds rapidly to symptomatic treatment (eg, antihistamines, NSAIDs, narcotic agents, intravenous fluids); requires ≤ 24 hours of prophylactic dosing Grade III Delay (eg, no prompt response to symptomatic treatment and/or discontinuation of instillation over time); relapse after improvement; hospitalization required for deuteropathy (eg, renal impairment, pulmonary infiltrates) Grade IV Life threatening; positive pressure breathing or artificial respiration required Grade 5 (Grade V) die

[Table 7] (CTCAE v 5.0) Grade I Fever with or without systemic symptoms Grade II Hypotension responsive to infusion; hypoxia responsive to <40% oxygen Grade III Hypotension controllable with a single dose of vasopressors; hypoxia requiring ≥40% oxygen Grade IV Life threatening; urgent action required Grade 5 (Grade V) die

[Table 8] (Lee's Criteria) Grade I Not life-threatening; only symptomatic treatment required (fever, nausea, fatigue, headache, muscle pain, malaise) Grade II Appropriate intervention required; hypoxia responsive to >40% oxygen administration; hypotension responsive to single-dose IV fluids or low-dose vasopressors; grade 2 or higher organ toxicity Grade III Active intervention required; hypoxia responsive to ≤40% oxygen; hypotension responsive to high doses or multiple doses of vasopressors; organ toxicity of grade 3 or higher or high blood pressure of grade 4 or higher transaminemia Grade IV Life-threatening; artificial respiration required; organ toxicity of grade 4 or higher (except hypertransaminemia) Grade 5 (Grade V) die

[Table 9] (ASTCT CRS Consensus Grading) fever Hypotension and/or hypoxia Grade I Above 38℃ as well as Hypotension: none, hypoxia: none Grade II Above 38℃ as well as Hypotension not requiring vasopressors, and/or hypoxia requiring low-flow nasal cannula or blow-by Grade III Above 38℃ as well as Hypotension requiring a single dose of a vasopressor (with or without concomitant vasopressin) and/or hypoxia requiring a high-flow nasal cannula, mask, non-rebreathing mask, or Venturi mask Grade IV Above 38℃ as well as Hypotension requiring multiple vasopressors (minus vasopressin), and/or hypoxemia requiring positive pressure (eg, CPAP, BiPAP, intubation, and mechanical ventilation)

In one aspect, the symptoms of CRS occur within minutes, hours, or days after the start of administration of the drug, and some are regarded as late-onset symptoms. Preferably the symptoms of CRS occur within about 96 hours, within about 72 hours, or within about 48 hours after the start of administration of the anti-T cell antigen-binding molecule, more preferably from the administration of the anti-T cell antigen-binding molecule Occurs from the same day to the next day. In other aspects, the severity of symptoms of CRS correlates with peak cytokine concentrations.

Signs of CRS In one aspect, the signs of CRS refer to the above-mentioned signs of CRS (regardless of grade), similar to the symptoms of CRS. Specific examples include, but are not limited to, fever or hypotension that occurs initially after administration of the anti-T cell antigen-binding molecule.

Treatment of CRS In one aspect, as a method for treating CRS, VEGF inhibitors, Bazedoxifene, SGP130 blockers, vasopressors, systemic corticosteroids (eg, corticosteroids), immunosuppressants, and mechanical artificial respiration can be mentioned. .

In one aspect, VEGF inhibition is selected from one or more of anti-VEGF antigen binding molecules, anti-VEGFR1 antigen binding molecules, anti-VEGFR2 antigen binding molecules, VEGF receptors or fusion proteins comprising fragments thereof, and tyrosine kinase inhibitors agent or antagonist inhibitor. Preferred VEGF inhibitors are disclosed in "III. VEGF inhibitors" in this specification.

In one aspect, when the VEGF inhibitor is bevacizumab, the active ingredient is about 5 to 15 mg/kg per dose, for example, 5 mg/kg or less per dose can be selected for patients with a body weight of 30 kg or more. amount, 7.5 mg/kg or less, 10 mg/kg or less, 15 mg/kg or less, for example, over 0.5 hours, 1 hour, 2 hours, or 3 hours, via Administered by intravenous injection. In one aspect, when the VEGF inhibitor is ramucirumab, the active ingredient is about 4 to 15 mg/kg each time, for example, for a patient with a body weight of 30 Kg or more, 8 mg/kg or The amounts below, 10 mg/kg or below, are administered via intravenous injection over, for example, 0.5 hour, 1 hour, 2 hours, or 3 hours. In different aspects, bevacizumab (Avastin) and ranibizumab (Lucentis) are administered for the purpose of treatment of CRS. When the symptoms of CRS are not improved after the first administration, the most Three additional doses of bevacizumab (Avastin) and ranibizumab (Lucentis) were administered. In one example, in the case of continuous administration of bevacizumab (Avastin) and ranibizumab (Lucentis), an interval of 8 hours or more is required from the previous administration.

Exemplary vasoactive agents are not limited to these, but angiotensin-11 (angiotensin-11), endothelin-1 (endothelin-1), and alpha-adrenergic agonist can be mentioned. , prostanoids, phosphodiesterase inhibitors, endothelin antagonists, circulatory enhancers (eg, epinephrine, dobutamine, isoprenaline, ephedrine), Blood pressure agents (eg, norepinephrine, vasopressin, metaraminol, vasopressin, methylene blue), cardiotonic vasodilators (eg, milrinone, levosimendan), and dopamine .

Examples of blood pressure raising agents include, but are not limited to, norepinephrine, dopamine, phenylephrine, epinephrine, and vasopressin. In one aspect, the blood pressure raising agent comprises a high dose blood pressure raising agent and a low dose blood pressure raising agent. In one aspect, the high dose blood pressure raising agent is the following: a monotherapy comprising norepinephrine above 20 μg/min, a single dose of dopamine above 10 μg/kg/min, adrenaline above 200 μg/min One or more of single-dose therapy and/or epinephrine single-dose therapy of 10 μg/min or more. In some embodiments, when the individual is being administered vasopressin, the high-dose blood pressure raising agent contains vasopressin + norepinephrine equivalent of 10 μg/min or more (here, norepinephrine and the like). Dose of effector = [norepinephrine (μg/min)] + [dopamine (μg/kg/min)/2] + [epinephrine (μg/min)] + [dehydrocarbon epinephrine (μg/min)/ 10]). In some embodiments, when the individual is being administered a combined blood pressure-raising agent (other than vasopressin), the high-dose blood pressure-raising agent contains 20 μg/min or more of a norepinephrine equivalent (here, the Norepinephrine equivalent dose = [norepinephrine (μg/min)] + [dopamine (μg/kg/min)/2] + [epinephrine (μg/min)] + [dehydrocarbon epinephrine ( μg/min)/10]). For example, see above.

In one aspect, a low-dose blood pressure-raising agent is a high-dose blood pressure-raising agent administered in a dose smaller than the one or more doses listed above.

Exemplary corticosteroids include, but are not limited to, dexamethasone, hydrocortisone, and methylprednisolone. In one aspect, dexamethasone is used at a dose of 0.5 mg/kg via oral or intravenous administration. In one aspect, 8-20 mg of dexamethasone, a pharmaceutically acceptable salt thereof, or a derivative thereof per dose is used via oral or intravenous administration. It should be understood by those skilled in the art that the dose of dexamethasone to be administered is not limited to the above, and can be appropriately changed according to the state of the individual and the severity of CRS.

Examples of immunosuppressive agents include TNF-α inhibitors or IL-1 inhibitors. In one aspect, the inhibitor of TNF-alpha comprises an anti-TNF-alpha antibody, eg, a monoclonal antibody, eg, Infliximab. In one aspect, the inhibitor of TNF-alpha comprises a soluble TNF-alpha receptor (eg, tumor necrosis factor receptor fusion protein (Etanercept)). In one embodiment, the IL-1 or IL-1R inhibitor comprises Anakinra.

V. Combination therapy In one aspect, the present disclosure pertains to a concomitant use of any agent that is responsible for either or both of CRS and cytokine release in a subject to be administered, with a VEGF inhibitor , for any one or a combination thereof selected for the purpose of preventing, reducing, and treating CRS and cytokines resulting from the administration of the agent Release either or both. Combination therapy in the present disclosure can be defined as follows: That is, the combination therapy is the combination therapy of the agent A and the agent B. The agent A has the effect of inducing either or both of CRS and cytokine release by administration to the subject, and the agent B is a VEGF inhibitor. Selected from any one, or a combination thereof, for the purpose of preventing, reducing, and treating, the prevention, alleviation, and treatment being the prevention, alleviation, and treatment of CRS and cytokine release induced by the administration of Agent A either or both. In yet another aspect, the present disclosure relates to a pharmaceutical composition comprising an arbitrary agent (agent A) for use in combination therapy with a VEGF inhibitor (agent B), wherein the aforementioned agent is Administer an agent that induces either or both of CRS and cytokine release in a subject, for a pharmaceutical composition selected from any one or a combination thereof for the purpose of prevention, mitigation, and treatment, the prevention, Mitigation, and treatment is the prevention, mitigation, and treatment of either or both of CRS and cytokine release resulting from administration of the agent. In another aspect, the present disclosure relates to a pharmaceutical composition comprising a VEGF inhibitor for use in combination therapy with any agent that induces CRS and cells in a subject to be administered A pharmaceutical composition of either or both of hormone release, for use in a pharmaceutical composition selected from any one of, or a combination of, the purpose of prevention, alleviation, and treatment, the prevention, alleviation, and treatment being prevention, alleviation , and treating either or both of CRS and cytokine release resulting from administration of the agent. In the present disclosure, in the case where the agent that has the potential to induce either or both of CRS and cytokine release is an anti-T cell antigen-binding molecule, it relates to a method for treating cancer, which is characterized by inhibiting the release of VEGF through VEGF Administration of an agent selected from any one of, or a combination of, the purpose of preventing, alleviating, and treating, preventing, alleviating, and treating CRS and Either or both of cytokine release. In the present disclosure, an agent capable of inducing either or both of CRS and cytokine release in an administration subject means that there is a possibility of causing either or both of CRS and cytokine release. Pharmacy. Specifically, agents that have been reported to induce either or both of CRS and cytokine release are included as side effects accompanying administration. For example, a drug targeting lymphocytes is a drug that has the potential to induce cytokine production due to its stimulation. In other words, agents with lymphocyte stimulatory properties may be responsible for either or both of CRS and cytokine release. As an example of such a drug, a drug containing an anti-T cell antigen-binding molecule as an active ingredient can be exemplified.

In one aspect, by administration of a VEGF inhibitor, in an individual, the selection is made from any one of, or a combination of, prevention, amelioration, and treatment for the purpose of prevention, amelioration, and treatment. and treatment of either or both of CRS and cytokine release syndrome. Thus, the present disclosure can also be stated in other words as a method comprising administering a VEGF inhibitor for any one, or a combination thereof, selected for the purpose of preventing, alleviating, and treating, the preventing, alleviating, and treating being prophylactic, Alleviate, and treat the occurrence of either or both of CRS and cytokine release.

In one aspect, the VEGF inhibitor is administered subcutaneously or intravenously to the individual prior to, concurrently with, or after administration of any agent responsible for either or both of CRS and cytokine release. Preferably, the VEGF inhibitor is administered intravenously to the individual prior to, concurrently with, or after administration of the aforementioned agents. Not intended to be limiting, when a VEGF inhibitor is administered before or at the same time as the aforementioned agent, it has the ability to prevent or reduce the occurrence of either or both of CRS and cytokine release accompanying the administration of the agent. Effect. In one aspect, the VEGF inhibitor is administered 6 days, 5 days, 4 days, 3 days, 2 days before administration of any agent that causes either or both of CRS and cytokine release , 1 day before or on the same day, before administration of the drug. Alternatively, the VEGF inhibitor can also be administered on the day of administration of the agent at the same time as the agent (in the present disclosure, before the administration of any agent that will cause either or both of CRS and cytokine release The administration of the VEGF inhibitor and the administration of the VEGF inhibitor concurrently with the administration of the drug are collectively referred to as "pre-administration of the VEGF inhibitor"). In one aspect, pre-administration of a VEGF inhibitor does not significantly impair the efficacy of any agent responsible for either or both of CRS and cytokine release (eg, TDCC (T cell-dependent cellular). cytotoxicity; T cell-dependent cytotoxic activity), antitumor effect, etc.), and prevent or reduce the occurrence of CRS accompanying the administration of the agent.

Furthermore, when a VEGF inhibitor is administered after administration of any drug that causes either or both of CRS and cytokine release, or before the occurrence of CRS, it has the potential to prevent or alleviate CRS that may occur later. Effect. Furthermore, when the VEGF inhibitor is administered after the administration of the drug or after the occurrence of CRS or symptoms of CRS, it has the effect of treating CRS and alleviating the symptoms.

Accordingly, the present disclosure provides a method for either or both of the treatment of CRS and the alleviation of symptoms, comprising the step of administering a VEGF inhibitor to a subject selected for either or both of CRS and cytokine release or Arbitrary medical treatment for both causes, monitoring for the occurrence of CRS for signs of it. Methods for monitoring the occurrence of CRS for signs of it are well known as previously described.

The effects of the selection of VEGF inhibitors from any one of or a combination for the purpose of preventing, alleviating, and treating CRS are the first clear effects in this disclosure , and either, or both, of cytokine release. Accordingly, the present disclosure provides a pharmaceutical composition, including a VEGF inhibitor, for use in any one or a combination selected for the purpose of prevention, alleviation, and treatment, the prevention, alleviation, and treatment being Either or both of CRS and cytokine release are prevented, alleviated, and treated. Alternatively, the present disclosure provides the use of a VEGF inhibitor in the preparation of a pharmaceutical composition for the purpose selected from either or both of the prevention, mitigation, and treatment of CRS and cytokine release. one, or a combination thereof. Further, the present disclosure provides the use of a VEGF inhibitor for any one or a combination thereof selected for the purpose of prevention, alleviation, and treatment, the prevention, alleviation, and treatment being the prevention, alleviation, and treatment of CRS and cytokines Either or both of the releases. Furthermore, the present disclosure provides a method comprising the steps of, selecting a subject selected from any one of the following, and administering a VEGF inhibitor: a subject developing symptoms of either or both of CRS and cytokine release, observing the The object of the occurrence of the symptom, and the object of the treatment with the possibility of either or both of CRS and cytokine release, are selected for the purpose of any one of prevention, mitigation, and treatment, or In combination, the prevention, alleviation, and treatment is either or both of preventing, alleviating, and treating CRS and cytokine release. Furthermore, in the present disclosure, treatment that may be accompanied by either or both of CRS and cytokine release includes, for example, administration of an anti-T cell antigen-binding molecule.

In one aspect, the anti-T cell antigen-binding molecule used in the combination therapy comprises "a domain comprising an antibody variable region having T cell receptor complex-binding activity" and "comprising an antibody having cancer antigen-binding activity" The bispecific antigen-binding molecule of the variable region "domain" is preferably a bispecific antibody. In one aspect, the bispecific antibody may also be in the form of a single-chain antibody, eg, in which the variable regions of the antibody are linked by a linker. In one aspect, the anti-T cell antigen-binding molecule further comprises an Fc region with reduced binding activity to Fcγ receptors. Preferably, the anti-T cell antigen-binding molecule comprises (1) a domain comprising an antibody variable region having Glypican3 binding activity, (2) a domain comprising an antibody variable region having T cell receptor complex binding activity, and ( 3) A bispecific antibody comprising a domain of an Fc region whose binding activity to an Fcγ receptor is reduced.

In one aspect, the VEGF inhibitor used in the combination therapy is a VEGF inhibitor, preferably an antibody against VEGF or its receptor, most preferably bevacizumab or ramucirumab. In another aspect, the VEGF inhibitor used in the combination therapy is a fusion protein containing a ligand-binding site of a VEGF receptor, and aflibercept can be mentioned as a preferred example. In one aspect, when bevacizumab and ramucirumab are administered before or at the same time as the anti-T cell antigen-binding molecule, bevacizumab and ramucirumab are effective in human adults and adults. For example, from 5mg/kg to 100mg/kg, for example from 5mg/kg to 90mg/kg, 5mg/kg to 80mg/kg, 5mg/kg to 70mg/kg, 5mg/kg to 60mg/kg, 5mg/kg ~50mg/kg, 5mg/kg~40mg/kg, 5mg/kg~30mg/kg, 5mg/kg~20mg/kg, 5mg/kg~15mg/kg, or for example, 10mg/kg~100mg/kg, 20mg/kg kg~100mg/kg, 30mg/kg~100mg/kg, 40mg/kg~100mg/kg, 50mg/kg~100mg/kg, 60mg/kg~100mg/kg, 70mg/kg~100mg/kg, 80mg/kg~ A dose selected from among 100 mg/kg, 90 mg/kg to 100 mg/kg, etc., is administered as a single dose. Also, for the avoidance of doubt, for example, when it is described as 5 mg to 100 mg/kg, the same as 5.0 mg/kg, 5.1 mg/kg, 5.2 mg/kg, 5.3 mg/kg, 5.4 mg/kg, 49.9 mg /kg, 50mg/kg, 50.1mg/kg, 50.2mg/kg,... All doses between /kg are individually and specifically described in this specification. In one aspect, before or at the same time as the administration of the anti-T cell antigen-binding molecule, for patients with a body weight of 30 kg or more, each administration is 15 mg/kg or less, 0.5-15 mg/kg, 1-15 mg/kg, 2-15 mg/kg. 15mg/kg, 3~15mg/kg, 4~15mg/kg of bevacizumab. In one aspect, 10 mg/kg or less, 0.5 to 10 mg/kg, 1 to 10 mg/kg, 2 to 10 mg/kg, 0.5 to 10 mg/kg, 1 to 10 mg/kg, 2 to 10mg/kg, 3~10mg/kg, 4~10mg/kg of ramucirumab.

According to the present disclosure, in some embodiments, in addition to the combined use of the aforementioned anti-T cell antigen-binding molecule and a VEGF inhibitor, an immune checkpoint inhibitor can also be used in combination. "Immune checkpoint" refers to a molecule that is expressed on immune-responsible cells (including T cells), and through binding to a ligand, transmits a message to inhibit an immune response to the immune-responsible cell. Immune checkpoints and their ligands include, for example, PD-1, CTLA-4, TIM3, LAG3, PD-L1, PD-L2, BTNL2, B7-H3, B7-H4, CD48, CD80, 2B4, BTLA, CD160, CD60 , CD86 or VISTA, etc., but not limited thereto. In some embodiments, the "immune checkpoint inhibitor" in the present invention refers to an agent that inhibits the signal transduction through an immune checkpoint by inhibiting the binding of an immune checkpoint to its ligand.

As a non-limiting aspect of the present disclosure, preferred examples of immune checkpoint inhibitors include PD1 antibody, PDL1 antibody, CTLA-4 antibody, TIM3 antibody, or LAG3 antibody, but are not limited thereto. For example, examples of PD-1 antibodies include Pembrolizumab (CAS Accession No.: 1374853-91-4), Nivolumab (CAS Accession No.: 946414-94-4), MEDI0680, PDR001, BGB-A317, REGN2810, SHR- 1210, PF-06801591 and other well-known PD1 antibodies. Examples of PD-L1 antibodies include atezolizumab (CAS accession number: 1380723-44-3), Avelumab (CAS accession number: 1537032-82-8), and Durvalumab (CAS accession number: 1428935-60) -7), MDX-1105 and other well-known PD-L1 antibodies. Examples of CTLA-4 antibodies include various known CTLA-4 antibodies such as Ipilimumab (CAS Accession No.: 477202-00-9) and Tremelimumab (CAS Accession No.: 745013-59-6). Examples of the TIM3 antibody include various well-known TIM3 antibodies such as MBG452. Examples of the LAG3 antibody include various well-known LAG3 antibodies such as BMS-986016 and LAG525. In yet another aspect of the present disclosure, when an immune checkpoint inhibitor is additionally used in combination with the aforementioned anti-T cell antigen-binding molecule and a VEGF inhibitor, the immune checkpoint inhibitor can be administered at any time point to the object. Specifically, for example, the immune checkpoint inhibitor can be administered before or after the combined use of the anti-T cell antigen-binding molecule and the VEGF inhibitor. Therefore, for example, when the anti-T cell antigen-binding molecule and the VEGF inhibitor or the immune checkpoint inhibitor are used in combination to evaluate the cancer regression effect, the other can be additionally administered. Alternatively, the above-mentioned three doses can be administered simultaneously.

The concomitant therapy of the present disclosure may further comprise administration of corticosteroids. In one aspect, for the purpose of preventing or reducing CRS accompanying the administration of the anti-T cell antigen-binding molecule, 2 days before, 1 day before, or 0 days before the day of administration of the anti-T cell antigen-binding molecule The day before (the same day), before the administration of the anti-T cell antigen binding molecule, the corticosteroid is administered orally or intravenously to the individual. Alternatively, concurrently with administration of the anti-T cell antigen-binding molecule, corticosteroids are administered to the individual orally or intravenously (these are referred to as "pre-administration of corticosteroids" or "pre-administration of steroids"). The pre-administration of the corticosteroid is preferably performed in addition to the pre-administration of the VEGF inhibitor (in combination with the pre-administration of the VEGF inhibitor). It is not intended to be limiting, and examples of preferable corticosteroids include dexamethasone, hydrocortisone, and methylprednisolone. The corticosteroid is preferably dexamethasone, a pharmaceutically acceptable salt thereof, or a derivative thereof, administered via oral or intravenous administration. It should be understood by those skilled in the art to which the invention pertains that the dose of dexamethasone is not limited to the above, and can be appropriately changed according to the state of the individual, the occurrence of CRS, and the like.

In one aspect, the combination therapy follows (1) pre-administration of corticosteroids, (2) pre-administration of VEGF inhibitor, (3) anti-T cell antigen-binding molecule on the day of administration of the anti-T cell antigen-binding molecule order of administration. In this case, for example, 1 hour or more, 2 hours or more, 3 hours or more, 4 hours or more, or 5 hours or more before the start of administration of the VEGF inhibitor, preferably 2 hours or more before, the individual who stops the corticosteroid VEGF inhibition is terminated more than 1 hour, more than 2 hours, more than 3 hours, more than 4 hours, or more than 5 hours, preferably more than 2 hours before the start of administration of the anti-T cell antigen-binding molecule Administration of an agent to an individual. In another aspect, the administration on the day of administration of the anti-T cell antigen-binding molecule is performed with (1) pre-administration of VEGF inhibitor, (2) pre-administration of corticosteroid, (3) anti-T cell antigen binding The order of administration of the molecules is performed. In this case, for example, 1 hour or more, 2 hours or more, 3 hours or more, 4 hours or more, or 5 hours or more, preferably 2 hours or more before the start of administration of the corticosteroid, for the individual who stops the VEGF inhibitor 1 hour, 2 hours or more, 3 hours or more, 4 hours or more, or 5 hours or more, preferably 2 hours or more before the start of administration of the anti-T cell antigen-binding molecule, the corticosteroids are terminated the contribution to the individual.

In a different aspect, the concomitant therapy of the present disclosure does not comprise pre-administration of a corticosteroid. Not for limitation, prevention or reduction of the occurrence of CRS accompanying administration of anti-T cell antigen-binding molecule by pre-administration of VEGF inhibitor does not require administration of corticosteroids for the purpose of prevention or reduction of CRS. give.

The concomitant therapy of the present disclosure may further comprise administration of Tocilizumab. In one aspect, for the purpose of preventing or reducing CRS accompanying the administration of the anti-T cell antigen-binding molecule, 2 days before, 1 day before, or 0 days before the day of administration of the anti-T cell antigen-binding molecule The day before (the day), prior to the administration of the anti-T cell antigen-binding molecule, tocilizumab was administered intravenously to the individual. Alternatively, concurrently with the administration of the anti-T cell antigen-binding molecule, tocilizumab is administered intravenously to the individual (these are referred to as "pre-administration of tocilizumab" or "pre-administration of tocilizumab"). ".) The pre-administration of tocilizumab is preferably performed in addition to the pre-administration of the VEGF inhibitor (in combination with the pre-administration of the VEGF inhibitor). Those with ordinary knowledge in the technical field to which the invention pertains should understand that the dose of tocilizumab is not limited to the above, and can be appropriately changed according to the state of the individual, the occurrence of CRS, and the like.

In one aspect, concomitant therapy is based on (1) pre-administration of tocilizumab, (2) pre-administration of VEGF inhibitor, (3) anti-T cell pre-administration on the day of administration of the anti-T cell antigen-binding molecule The order of administration of the antigen-binding molecule is performed. In this case, for example, 1 hour or more, 2 hours or more, 3 hours or more, 4 hours or more, or 5 hours or more before the start of administration of the VEGF inhibitor, preferably 2 hours or more, tocilizumab is terminated The administration of the anti-T cell antigen-binding molecule to the individual, preferably 2 hours or more before the start of administration of the anti-T cell antigen-binding molecule , to terminate the administration of the VEGF inhibitor to the individual. In another aspect, the administration of the anti-T cell antigen-binding molecule on the day of administration is performed with (1) pre-administration of VEGF inhibitor, (2) pre-administration of tocilizumab, (3) anti-T cell The order of administration of the antigen-binding molecule is performed. In this case, for example, the VEGF inhibitor is terminated 1 hour or more, 2 hours or more, 3 hours or more, 4 hours or more, or 5 hours or more, preferably 2 hours or more before the start of tocilizumab administration. and more than 1 hour, more than 2 hours, more than 3 hours, more than 4 hours, or more than 5 hours before the administration of the anti-T cell antigen-binding molecule, preferably more than 2 hours before, The administration of tocilizumab to the individual is discontinued.

In a different aspect, the concomitant therapy of the present disclosure does not comprise pre-administration of tocilizumab. It is not intended to be limiting. Tocilizumab is not required for the purpose of preventing or reducing CRS by pre-administration of a VEGF inhibitor to prevent or reduce the occurrence of CRS accompanying the administration of an anti-T cell antigen-binding molecule. 's investment.

With regard to any of the anti-T cell antigen binding molecules, VEGF inhibitors, and/or other agents in the present disclosure, those of ordinary skill in the art to which the invention pertains will of course understand the timing of administration, interval of administration, and administration Each quantity can have a certain "tolerable range", and a person with ordinary knowledge in the technical field to which the invention pertains can appropriately determine the tolerable range. For example, based on the individual's symptoms, etc., and at the discretion of the physician, the administration interval of the anti-T cell antigen-binding molecule, the VEGF inhibitor, and/or other drugs may be appropriately increased or decreased, or the dose of the drug may be appropriately increased or decreased. within the "tolerable range".

Non-limiting specific examples of combined therapy comprising an anti-T cell antigen-binding molecule and a VEGF inhibitor are described below.

Administration of Anti-T Cell Antigen Binding Molecules with Pre-Administration of VEGF Inhibitor In one aspect, the present disclosure provides a pharmaceutical composition comprising an anti-T cell antigen binding molecule for use in combination therapy with a VEGF inhibitor. In yet another aspect, the present disclosure provides a pharmaceutical composition comprising a VEGF inhibitor for use in combination therapy with an anti-T cell antigen binding molecule. In a further aspect, the VEGF inhibitor is administered to the individual prior to or concurrently with the administration of the aforementioned anti-T cell antigen-binding molecule. Without limitation, the VEGF inhibitor has an effect of preventing or reducing the occurrence of CRS accompanying the administration of the anti-T cell antigen-binding molecule when administered before or simultaneously with the administration of the anti-T cell antigen-binding molecule. In one aspect, the VEGF inhibitor is administered 6 days, 5 days, 4 days, 3 days, 2 days, 1 day or the day before the administration of the anti-T cell antigen binding molecule, on the aforementioned bispecific. Administration of the antibody is performed prior to administration. Alternatively, the VEGF inhibitor is administered simultaneously with the aforementioned anti-T cell antigen-binding molecule on the day of administration of the anti-T cell antigen-binding molecule. In one aspect, the efficacy of anti-T cell antigen binding molecules (eg, TDCC (T cell-dependent cellular cytotoxicity; T cell-dependent cellular cytotoxicity), anti-tumor effect, etc.), and prevent or reduce the occurrence of CRS accompanying the administration of the anti-T cell antigen-binding molecule.

VI. Combination therapy of bispecific antibody against Glypican3 (GPC3)/T cell receptor complex and VEGF inhibitor In one aspect, the present disclosure provides a bispecific antibody comprising an anti-GPC3/T cell receptor complex for use in a pharmaceutical composition for concomitant therapy with a VEGF inhibitor. In yet another aspect, the present disclosure provides a pharmaceutical composition comprising a VEGF inhibitor for use in combination therapy with an anti-Glypican3 (GPC3)/T cell receptor complex bispecific antibody. In one aspect, the VEGF inhibitor is administered to the individual prior to, or concurrently with, administration of the aforementioned bispecific antibody to the GPC3/T cell receptor complex.

Not intended to be limiting, when the VEGF inhibitor is administered before or simultaneously with the administration of the anti-GPC3/T cell receptor complex bispecific antibody, it has the effect of preventing or reducing the risk associated with the anti-GPC3/T cell receptor complex. The effect of CRS on administration of complex bispecific antibodies. In one aspect, the VEGF inhibitor is 6 days, 5 days, 4 days, 3 days, 2 days, 1 day before administration of the anti-GPC3/T cell receptor complex bispecific antibody Before or on the day, before administration of the aforementioned bispecific antibody. Alternatively, the VEGF inhibitor is administered simultaneously with the aforementioned bispecific antibody on the day of administration of the anti-GPC3/T cell receptor complex bispecific antibody (pre-administration of the VEGF inhibitor). As a specific example of the combination therapy including the pre-administration of a VEGF inhibitor, please refer to the above-mentioned various aspects. In one aspect, the efficacy of the bispecific antibody against the GPC3/T cell receptor complex (eg, TDCC (T cell-dependent cellular cytotoxicity; T cell) is not significantly impaired by pre-administration of the VEGF inhibitor. cell-dependent cytotoxic activity), antitumor effect, etc.), preventing or reducing the occurrence of CRS accompanying the administration of the anti-GPC3/T cell receptor complex bispecific antibody.

VII. Manufacturing method and kit Furthermore, the present invention provides a kit for use in the method of the present invention, comprising the anti-T cell antigen-binding molecule of the present invention or an anti-T cell antigen-binding molecule produced by the production method of the present invention. Other pharmaceutically acceptable carriers, media, instructions for use, and the like can also be included in the kit. Furthermore, the present invention relates to an anti-T cell antigen-binding molecule of the present invention used in the method of the present invention or an anti-T cell antigen-binding molecule produced by the production method of the present invention. Furthermore, the present invention provides kits for use in the methods of the present invention, comprising VEGF inhibitors. The kit can also be packaged in advance with other pharmaceutically acceptable carriers, media, instructions for recording the usage method, and the like. The present invention is also directed to VEGF inhibitors for use in the methods of the present invention.

Furthermore, the present invention also relates to nucleic acids encoding these molecules, vectors into which the nucleic acids are introduced, cells comprising the nucleic acids or the vectors, methods of culturing the cells, producing the molecules, and produced by the methods. Anti-T cell antigen binding molecules or VEGF inhibitors.

For example, the anti-T cell antigen-binding molecule can be produced by appropriately introducing a nucleic acid encoding an anti-T cell antigen-binding molecule or a vector into which the nucleic acid is introduced into a host cell, followed by culturing the host cell. When the anti-T cell antigen-binding molecule is secreted, the target anti-T cell antigen-binding molecule can be recovered from the culture supernatant. Methods for purifying a desired anti-T cell antigen-binding molecule from culture are also known. On the other hand, the same goes for VEGF inhibitors. If they are protein components such as antibodies, a nucleic acid encoding the VEGF inhibitor molecule or a vector into which the nucleic acid is introduced is appropriately introduced into a host cell, and then the host cell can be cultured to interact with the VEGF inhibitor. The multispecific antigen-binding molecule was produced in the same way.

In the present disclosure, the anti-T cell antigen-binding molecule and the VEGF inhibitor can be individually packaged together with other pharmaceutically acceptable carriers, vehicles, and instructions describing the method of use. Or after formulating both, it becomes a combination kit. In particular, the kit may also comprise a package containing a formulated anti-T cell antigen binding molecule, and a package containing a formulated VEGF inhibitor. When each formulation constituting the kit is a freeze-dried or liquid concentrated formulation, the kit can also be combined with a liquid medium for dissolving or diluting the formulation.

The present disclosure provides a pharmaceutical composition comprising the above-mentioned anti-T cell antigen-binding molecule, which is used for either or both of the treatment and prevention of cancer co-administered with a VEGF inhibitor A pharmaceutical composition for any one or a combination thereof selected for the purpose of prevention, relief, and treatment resulting from the administration of the aforementioned pharmaceutical composition Either or both of CRS and cytokine release are administered.

Here, the above-mentioned pharmaceutical composition is not limited to a pharmaceutical composition containing an anti-T cell antigen-binding molecule, and may be a general peripheral gene in which a gene encoding a chimeric antigen receptor (Chimeric Antigen Receptor: hereinafter, referred to as "CAR") is introduced. CAR-expressing T cells (hereinafter, referred to as "CAR-T cells") produced from blood T cells (peripheral blood T lymphocytes). CARs artificially synthesize antibodies that recognize cell surface antigens such as cancer cells, and chimeric proteins that induce signaling regions for activation of T cells. CAR-T cells are produced by introducing peripheral blood T lymphocytes into a gene encoding CAR. The CAR-expressing T cells produced by this method are used for the treatment of cancer diseases by adoptive immunotherapy. Such CAR-T cells are reactive against antigen-expressing target cells and can cause damage to target cells independent of interaction with the major histocompatibility complex (MHC).

Cancer immunotherapy through the administration of CAR-T cells, more specifically, is a therapy in which T cells are taken from a patient, a gene encoding a CAR is introduced into the T cells, and the T cells are expanded and cultured, and then transferred to the patient again. Clinical trials (Non-Patent Document 1). Cancer immunotherapy through the administration of CAR-T cells has shown results of efficacy against hematopoietic malignancies such as leukemia and lymphoma. In 2017, Kymriah (registered trademark) (Novartis, tisagenlecleucel, CTL-019, CD3 zeta-CD137) and Yescarta (registered trademark) (KiTE, axicabtagene ciloleucel, CD3 zeta-CD28) of CAR-T with CD19 as antigen have been Obtained a pharmaceutical license in the United States.

CRS and cytokine release resulting from cancer immunotherapy via administration of CAR-T cells have been reported (Lee DW, et al. Current concepts in the diagnosis and management of cytokine release syndrome. Blood. 2014 Jul 10; 124( 2): 188-95.). Since it has been shown that the CRS mechanism caused by the administration of a pharmaceutical composition containing an anti-T cell antigen-binding molecule and the administration of CAR-T cells are common, it is understood that the administration of a VEGF inhibitor can prevent the , alleviate, or treat cytokine release syndrome (CRS) accompanying the administration of CAR-T cells.

In addition, all the prior art documents cited in this specification are incorporated in this specification by reference. [Example]

[Example 1] Effect of concomitant use of anti-mouse VEGFR2 antibody (DC101) on the efficacy of ERY974 surrogate antibody (GC33/2C11) In the test using a syngenic mouse model, the test for evaluating the effect on the efficacy of the ERY974 surrogate antibody through the combined use of an anti-mouse VEGFR2 antibody and DC101 (manufactured by Bio X Cell (US)) is as follows conduct. Also, as a substitute antibody for ERY974, GC33/2C11 (the heavy chain on the GPC3 side is the amino acid sequence of SEQ ID NO: 433, the light chain on the GPC3 side is the amino acid sequence of SEQ ID NO: 434, the heavy chain on the CD3 side is the amino acid sequence of SEQ ID NO: 434, and the heavy chain on the CD3 side was used. The amino acid sequence of SEQ ID NO: 435, the light chain on the CD3 side is the antibody of the amino acid sequence of SEQ ID NO: 436).

MC38/hGPC3 cancer cells that express human GPC3 in large quantities in MC38, a mouse cancer cell line, were transplanted subcutaneously into C57BL/6J mice. The transplant day is taken as day 0. The mice were grouped into 4 groups on Day 14 according to tumor size. For grouped mice, (1) vehicle, (2) GC33/2C11 (5 mg/kg), (3) anti-mouse VEGFR2 antibody (DC101) ( 10 mg/kg), and (4) GC33/2C11 (5 mg/kg) and anti-mouse VEGFR2 antibody (10 mg/kg).

As a result, the combined use of GC33/2C1 and the anti-mouse VEGFR2 antibody enhanced the efficacy of each antibody compared to a single dose. (figure 1)

[Example 2] Effects on tumor-derived cytokine production via concomitant use of anti-mouse VEGFR2 antibody (DC101) to ERY974 surrogate antibody (GC33/2C11) In an assay using a syngenic mouse model, an assay to evaluate the effect of concomitant use of an anti-mouse VEGFR2 antibody (DC101) on tumor tissue-derived cytokine production by the ERY974 surrogate antibody (GC33/2C11), Proceed as follows.

MC38/hGPC3 cancer cells that express human GPC3 in large quantities in MC38, a mouse cancer cell line, were transplanted subcutaneously into C57BL/6J mice. The transplant day is taken as day 0. The mice were grouped into 4 groups on Day 13 according to tumor size. For grouped mice, tail vein administration on day 13 was performed with (1) vehicle, (2) GC33/2C11 (5 mg/kg), (3) anti-mouse VEGFR2 antibody (10 mg/kg), and (4) ) GC33/2C11 (5 mg/kg) and anti-mouse VEGFR2 antibody (10 mg/kg). The blood of the mice was collected before administration and 2, 6, and 24 hours after antibody administration, and plasma components were collected, and the cytokines in the plasma were measured. Cytokines were measured using mouse cytokine Chemokine Magnetic Bead Panel 1 (Millipore) reagent and MAGPIXRxPONT4.2 system (Merck).

As a result, compared with the case where only GC33/2C11 was administered, the combined use group of GC33/2C11 and the anti-mouse VEGFR2 antibody was considered to have a tendency to reduce the production amount of tumor-derived cytokines. Therefore, it was shown that the production of tumor-derived cytokines due to ERY974 was reduced by concomitant use of an angiogenesis inhibitor ( FIGS. 2A to 2E ).

[Example 3] Effects of concomitant use of anti-mouse VEGFR2 antibody (DC101) on normal tissue-derived cytokine production by ERY974 surrogate antibody (GC33/2C11) In an assay using human GPC3 gene knock-in mice, the effect of the concomitant use of an anti-mouse VEGFR2 antibody (DC101) on normal tissue-derived cytokine production by the ERY974 surrogate antibody (GC33/2C11) was evaluated The test of influence was carried out as follows.

The human GPC3 gene expressing human GPC3 was implanted into a mouse expressing human GPC3 under the expression control of a mouse GPC3 promoter (WO2018/038046). On the other hand, it has been reported that mice and humans have GPC3-positive tissues in common. It is believed that there is a mild expression of GPC3 in the lung, commonly in humans and mice (Clin. Cancer. Res (2004) 10, 8630-40). Therefore, by measuring the amount of cytokines produced by administration of GC33/2C11 in the above-mentioned human GPC3 gene-transplanted mice, it is possible to predict and evaluate the mechanism of cytokine production derived from normal tissues when ERY974 is administered to humans.

The human GPC3 gene expressing human GPC3 was implanted into 4 groups of mice, and (1) vehicle, (2) GC33/2C11 (5 mg/kg), (3) anti-mouse VEGF2 antibody (10) were administered into the tail vein respectively. mg/kg), and (4) GC33/2C11 (5 mg/kg) with anti-mouse VEGFR2 antibody (10 mg/kg). The blood of the mice was collected before administration and 2 hours after antibody administration, and plasma components were collected, and the cytokines in the plasma were measured. Cytokines were measured using mouse cytokine Chemokine Magnetic Bead Panel 1 (Millipore) reagent and MAGPIXRxPONT4.2 system (Merck).

As a result, the combined use group of GC33/2C11 and the anti-mouse VEGFR2 antibody tended to reduce the production amount of cytokines derived from normal tissues compared to the case where only GC33/2C11 was administered. Therefore, it was shown that the production of normal tissue-derived cytokines by ERY974 can be reduced by using an angiogenesis inhibitor in combination ( FIG. 3 ). [Industrial Availability]

The present disclosure is useful for the prevention, alleviation, or treatment of side effects caused by cytokines following administration of an anti-T cell antigen-binding molecule or the like. Anti-T cell antigen-binding molecules are attracting attention as a cancer treatment method and the like. Thus, in one aspect, the present disclosure is useful for the treatment of cancer via anti-T cell antigen binding molecules.

none.

[ Fig. 1 ] A graph showing the effect of concomitant use of an anti-mouse VEGFR2 antibody (DC101) on the efficacy of the ERY974 surrogate antibody (GC33/2C11). In the figure, the vertical axis shows the tumor volume (mm ³ ), and the horizontal axis shows the number of days after tumor transplantation, taking the day of tumor transplantation as day 0. [ FIG. 2-A ] A graph showing the effect of concomitant use of anti-mouse VEGFR2 antibody (DC101) on tumor-derived cytokine (IL-6) production by ERY974 surrogate antibody (GC33/2C11). In the figure, the vertical axis shows the plasma concentration (pg/mL) of IL-6, and the horizontal axis shows the components administered to mice, and "combo" is used in combination. The graph shows the elapsed time after the administration of the active ingredient as 0. [ Fig. 2-B ] A graph showing the effect of concomitant use of an anti-mouse VEGFR2 antibody (DC101) on tumor-derived cytokine (TNF) production by an ERY974 surrogate antibody (GC33/2C11). In the figure, the vertical axis shows the plasma concentration of TNF (pg/mL), and the horizontal axis shows the components administered to the mice, and "combination" means that both are used together. The graph shows the elapsed time after the administration of the active ingredient as 0. [ Fig. 2-C ] A graph showing the effect of concomitant use of anti-mouse VEGFR2 antibody (DC101) on tumor-derived cytokine (IFN gamma) production by ERY974 surrogate antibody (GC33/2C11). In the figure, the vertical axis shows the plasma concentration (pg/mL) of IFNγ, and the horizontal axis shows the components administered to mice, and "combination" means that both are used together. The graph shows the elapsed time after the administration of the active ingredient as 0. [ FIG. 2-D ] A graph showing the effect of concomitant use of anti-mouse VEGFR2 antibody (DC101) on tumor-derived cytokine (CXCL9) production by ERY974 surrogate antibody (GC33/2C11). In the figure, the vertical axis shows the plasma concentration (pg/mL) of CXC ligand 9, and the horizontal axis shows the components administered to mice, and "combination" means that both are used together. The graph shows the elapsed time after the administration of the active ingredient as 0. [ FIG. 2-E ] A graph showing the effect of concomitant use of anti-mouse VEGFR2 antibody (DC101) on tumor-derived cytokine (CXCL10) production by ERY974 surrogate antibody (GC33/2C11). In the figure, the vertical axis shows the plasma concentration (pg/mL) of CXCL10, and the horizontal axis shows the components administered to mice, and "combination" means that both are used together. The graph shows the elapsed time after the administration of the active ingredient as 0. [ Fig. 3 ] A graph showing the effect of the combined use of an anti-mouse VEGFR2 antibody (DC101) on normal tissue-derived cytokine production by the ERY974 surrogate antibody (GC33/2C11). In the figure, the vertical axis shows the plasma levels of IL-6 (upper left), TNFα (upper right), IFNγ (middle left), IL-2 (middle right), CXCL9 (lower left) and CXCL10 (lower right) 2 hours after administration Medium concentration (pg/mL), and the horizontal axis shows the administered active ingredient. [ Fig. 4 ] A diagram showing various numbers of light chain variable regions and Kabat et al.

Claims (15)

A pharmaceutical composition comprising an anti-T cell antigen-binding molecule, characterized in that the aforementioned pharmaceutical composition is used in combination with a vascular epithelial cell growth factor (VEGF) inhibitor to prevent and/or alleviate and/or Treatment of cytokine release syndrome and/or cytokine release. The pharmaceutical composition according to claim 1, wherein the VEGF inhibitor is administered before or simultaneously with the administration of the pharmaceutical composition. The pharmaceutical composition according to claim 1 or 2, wherein 6 days before, 5 days before, 4 days before, 3 days before, 2 days before, 1 day before or on the day of the aforementioned pharmaceutical composition, in the aforementioned pharmaceutical composition Before administration of the drug, the aforementioned VEGF inhibitor is administered. The pharmaceutical composition according to any one of claims 1 to 3, wherein the aforementioned VEGF inhibitor is selected from anti-VEGF antigen-binding molecules, anti-VEGFR1 antigen-binding molecules, anti-VEGFR2 antigen-binding molecules, VEGF receptors, or fragments thereof The group consisting of fusion proteins, and tyrosine kinase inhibitors. The pharmaceutical composition according to any one of claims 1 to 4, wherein the aforementioned VEGF inhibitor is selected from Bevacizumab, Ramucirumab, and Aflibercept. formed group. The pharmaceutical composition according to any one of claims 1 to 5, wherein no corticosteroid is administered before or simultaneously with the administration of the pharmaceutical composition. The pharmaceutical composition according to any one of claims 1 to 5, wherein the corticosteroid is further administered before, simultaneously with or after the administration of the aforementioned pharmaceutical composition. The pharmaceutical composition according to claim 6 or 7, wherein the corticosteroid is Dexamethasone, a pharmaceutically acceptable salt thereof, or a derivative thereof. The pharmaceutical composition according to any one of claims 1 to 8, wherein the aforementioned anti-T cell antigen-binding molecule is a bispecific antigen-binding molecule comprising: (1) a domain comprising an antibody variable region having T cell receptor complex binding activity, and, (2) A domain comprising an antibody variable region having cancer antigen-binding activity. The pharmaceutical composition according to any one of claims 1 to 9, wherein the aforementioned anti-T cell antigen-binding molecule is a bispecific antibody comprising: (1) a domain comprising an antibody variable region having T cell receptor complex binding activity, (2) a domain comprising an antibody variable region having glypican3-binding activity, and (3) A domain comprising an Fc region whose binding activity to an Fcγ receptor is reduced. The pharmaceutical composition according to any one of claims 1 to 10 is used for the treatment of cancer. The pharmaceutical composition according to claim 11, wherein the combined use enhances the antitumor effect compared to the single dose of the pharmaceutical composition or the VEGF inhibitor. The pharmaceutical composition according to any one of claims 1 to 11, wherein the cytokine release syndrome is caused by the release of cytokines in either or both of the non-tumor tissue and the tumor tissue. The pharmaceutical composition according to claim 13, wherein the tumor tissue is a tumor tissue comprising GPC3 expressing cells. A VEGF inhibitor for preventing and/or alleviating and/or treating cytokine release syndrome and/or cytokine release.

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